首页 期刊 论文 学科刊群 资讯 加盟 关于
EN
image
Article(id=1238813313346359685, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1238813307784712441, articleNumber=null, orderNo=null, doi=10.13343/j.cnki.wsxb.20250750, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1759766400000, receivedDateStr=2025-10-07, revisedDate=null, revisedDateStr=null, acceptedDate=1765209600000, acceptedDateStr=2025-12-09, onlineDate=1773285709939, onlineDateStr=2026-03-12, pubDate=1772553600000, pubDateStr=2026-03-04, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1773285709939, onlineIssueDateStr=2026-03-12, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1773285709939, creator=13701087609, updateTime=1773285709939, updator=13701087609, issue=Issue{id=1238813307784712441, tenantId=1146029695717560320, journalId=1192105938417971205, year='2026', volume='66', issue='3', pageStart='961', pageEnd='1466', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1773285708614, creator=13701087609, updateTime=1773291912509, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1238839328915378858, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1238813307784712441, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1238839328915378859, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1238813307784712441, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=1167, endPage=1177, ext={EN=ArticleExt(id=1238813314172637628, articleId=1238813313346359685, tenantId=1146029695717560320, journalId=1192105938417971205, language=EN, title=Quorum sensing regulators control the expression of phosphodiesterase GepA in Vibrio parahaemolyticus, columnId=1192149543992045670, journalTitle=Acta Microbiologica Sinica, columnName=Research Article, runingTitle=null, highlight=null, articleAbstract=

Objective To investigate the transcriptional regulation of quorum sensing (QS) regulators AphA, ToxR, and QsvR on the expression of the phosphodiesterase (GepA) gene gepA in Vibrio parahaemolyticus. Methods Total RNAs were extracted from the wild type (WT) and the mutant strains of aphA, toxR, and qsvR. Quantitative real-time PCR (qPCR) was carried out to calculate the transcriptional variation of gepA between WT and mutant strains. The regulatory DNA region of gepA was cloned into the upstream region of promoterless luxCDABEreporter gene in the pBBRlux plasmid. The recombinant plasmid was respectively transferred into the WT and mutant strains. Luminescence assay was used to test the regulatory effect of QS regulators on the expression of gepA. The primer extension assay was employed to detect the transcription start site and the promoter activity of gepA. The effects of QS regulators on gepA were evaluated based on the abundance of primer extension products. The regulatory DNA region of gepA was cloned into the upstream region of lacZ in the pHRP309 plasmid. The LacZ recombinant plasmid was transformed into EC100 λpir harboring pBAD33 or PBAD33-qsvR. Two-plasmid LacZ reporter assay was conducted to investigate the regulatory effects of QS regulators on the transcription of gepA in EC100 λpir. The regulatory DNA region of gepA was amplified by PCR, and the His recombinant proteins of QS regulators were purified. The electrophoretic mobility shift assay (EMSA) was performed to investigate whether QS regulators directly regulated the expression of gepA. Results At low cell density, the qPCR results showed that expression of gepA in ΔaphA and ΔtoxR were significantly lower than that in WT, indicating that AphA and ToxR activated the transcription of gepA. The luminescence assay showed that the transcriptional activity of the promoter region of gepA in ΔaphA and ΔtoxR was significantly lower than that in WT, further indicating that AphA and ToxR promoted the transcription of gepA. The primer extension assay detected that the transcription start site of gepA was located at the A nucleotide 30 bp upstream of the start codon ATG, and its transcriptional activity was activated by AphA. The EMSA result indicated that His-AphA and His-ToxR were unable to bind the promoter DNA region of gepA. At high cell density, both the qPCR and primer extension assay indicated that QsvR inhibited the transcription of gepA. The EMSA result demonstrated that His-QsvR directly bound to the promoter DNA region of gepA. Two-plasmid lacZ reporter assay demonstrated that QsvR inhibited the transcriptional activity of the promoter region of gepA in EC100 λpir. Conclusion AphA and ToxR indirectly activate while QsvR directly inhibits the transcription of gepA. Therefore, the transcription level of gepA is higher at low cell density and significantly decreases at high cell density.

, correspAuthors=Miaomiao ZHANG, Renfei LU, authorNote=null, correspAuthorsNote=
*E-mail: LU Renfei,
ZHANG Miaomiao,
, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Chao LIU, Xue LI, Xi LUO, Yiquan ZHANG, Miaomiao ZHANG, Renfei LU), CN=ArticleExt(id=1238813316957655608, articleId=1238813313346359685, tenantId=1146029695717560320, journalId=1192105938417971205, language=CN, title=群体感应相关调控子对副溶血弧菌磷酸二酯酶GepA的表达调控, columnId=1192149544164012138, journalTitle=微生物学报, columnName=研究报告, runingTitle=null, highlight=null, articleAbstract=

目的 研究群体感应系统(quorum sensing, QS)相关调控子AphA、ToxR和QsvR对副溶血弧菌磷酸二酯酶GepA编码基因gepA表达的调控关系。 方法 提取野生株(wild type, WT)和aphA、toxR、qsvR突变株的总RNA，通过实时定量PCR (quantitative real-time PCR, qPCR)初步分析QS相关调控子对gepA的转录影响；将gepA上游调控区DNA序列克隆入pBBRlux质粒无启动子区的表达生物冷光基因luxCDABE上游，并将lux重组质粒分别导入WT和突变株中，采用lux报告基因融合实验进一步研究QS相关调控子对gepA的调控作用；采用引物延伸(primer extension)法定位gepA的转录起始位点和核心启动子区，并根据引物延伸产物丰度判断QS相关调控子对gepA的调控关系；将gepA的调控区DNA序列克隆入pHRP309质粒中β-半乳糖苷酶基因上游获得LacZ重组质粒，将LacZ重组质粒分别转化入含有pBAD33或pBAD33-qsvR的大肠杆菌EC100 λpir中，采用LacZ报告基因融合实验研究在异体宿主中QS相关调控子对gepA转录的调控方式；PCR扩增gepA上游调控区DNA序列，同时表达并纯化QS相关调控子的His重组蛋白，采用凝胶阻滞实验(electrophoresis mobility shift assay, EMSA)研究QS相关调控子是否直接调控gepA的表达。 结果 低密度条件下，qPCR结果显示ΔaphA和ΔtoxR中gepA的转录水平明显低于WT，表明AphA和ToxR激活gepA的转录；lux报告基因融合实验显示，ΔaphA和ΔtoxR中gepA的启动子区转录活性明显低于WT，进一步表明AphA和ToxR促进gepA的转录；引物延伸结果显示，gepA的转录起始位点位于起始密码子ATG上游第30 bp的A，且其转录活性受到AphA的激活；EMSA结果显示，His-AphA和His-ToxR均不能与gepA的调控区DNA序列结合。高密度条件下，qPCR和引物延伸实验结果均显示QsvR抑制gepA的转录；EMSA实验结果显示，His-QsvR直接结合在gepA的启动子区DNA序列上；双质粒报告基因融合实验显示，QsvR可以抑制EC100 λpir中gepA的启动子区转录活性。 结论 AphA和ToxR间接激活，而QsvR直接抑制gepA的转录。因此，gepA在低密度条件下转录水平较高，高密度时转录水平明显下降。

, correspAuthors=张苗苗, 陆仁飞, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=ku81JSldbn8BZzWa0b7sog==, magXml=n7nONvzHg3WKhS2F8XUbmQ==, pdfUrl=null, pdf=X5Cx3uY3Fq86zGHcGSn7kg==, pdfFileSize=1285671, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=pSCrXuVuOCiE+QgtJPHKCw==, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=p6bQgwK0u/xMpIKO2aKLsA==, mapNumber=null, authorCompany=null, fund=null, authors=

作者贡献声明

刘超：提出研究思路、设计整体框架、获得关键实验数据；李雪：优化实验方案、承担实验操作；罗茜：验证结果可靠性；张义全：梳理研究现状，记录实验报告；张苗苗：分析实验数据、撰写文章并投稿；陆仁飞：提供基金支持，审阅文章，监督管理。

, authorsList=刘超, 李雪, 罗茜, 张义全, 张苗苗, 陆仁飞)}, authors=[Author(id=1238891100925121004, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, orderNo=0, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=0, email=null, emailSecond=null, emailThird=null, correspondingAuthor=0, authorType=1, ext={EN=AuthorExt(id=1238891101059338742, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, authorId=1238891100925121004, language=EN, stringName=Chao LIU, firstName=Chao, middleName=null, lastName=LIU, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=null, address=Department of Clinical Laboratory, Affiliated Nantong Hospital 3 of Nantong University, Nantong Third People’s Hospital, Nantong, Jiangsu, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1238891101180973571, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, authorId=1238891100925121004, language=CN, stringName=刘超, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=null, address=南通市第三人民医院，南通大学附属南通第三医院检验科，江苏 南通, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1238891100740571608, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, xref=null, ext=[AuthorCompanyExt(id=1238891100753154524, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, companyId=1238891100740571608, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Department of Clinical Laboratory, Affiliated Nantong Hospital 3 of Nantong University, Nantong Third People’s Hospital, Nantong, Jiangsu, China), AuthorCompanyExt(id=1238891100761543133, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, companyId=1238891100740571608, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=南通市第三人民医院，南通大学附属南通第三医院检验科，江苏 南通)])]), Author(id=1238891101327774220, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, orderNo=1, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=0, email=null, emailSecond=null, emailThird=null, correspondingAuthor=0, authorType=1, ext={EN=AuthorExt(id=1238891101466186266, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, authorId=1238891101327774220, language=EN, stringName=Xue LI, firstName=Xue, middleName=null, lastName=LI, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=null, address=Department of Clinical Laboratory, Affiliated Nantong Hospital 3 of Nantong University, Nantong Third People’s Hospital, Nantong, Jiangsu, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1238891101575238180, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, authorId=1238891101327774220, language=CN, stringName=李雪, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=null, address=南通市第三人民医院，南通大学附属南通第三医院检验科，江苏 南通, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1238891100740571608, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, xref=null, ext=[AuthorCompanyExt(id=1238891100753154524, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, companyId=1238891100740571608, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Department of Clinical Laboratory, Affiliated Nantong Hospital 3 of Nantong University, Nantong Third People’s Hospital, Nantong, Jiangsu, China), AuthorCompanyExt(id=1238891100761543133, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, companyId=1238891100740571608, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=南通市第三人民医院，南通大学附属南通第三医院检验科，江苏 南通)])]), Author(id=1238891101751398961, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, orderNo=2, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=0, email=null, emailSecond=null, emailThird=null, correspondingAuthor=0, authorType=1, ext={EN=AuthorExt(id=1238891101919171137, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, authorId=1238891101751398961, language=EN, stringName=Xi LUO, firstName=Xi, middleName=null, lastName=LUO, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=null, address=Department of Clinical Laboratory, Affiliated Nantong Hospital 3 of Nantong University, Nantong Third People’s Hospital, Nantong, Jiangsu, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1238891103441703499, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, authorId=1238891101751398961, language=CN, stringName=罗茜, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=null, address=南通市第三人民医院，南通大学附属南通第三医院检验科，江苏 南通, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1238891100740571608, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, xref=null, ext=[AuthorCompanyExt(id=1238891100753154524, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, companyId=1238891100740571608, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Department of Clinical Laboratory, Affiliated Nantong Hospital 3 of Nantong University, Nantong Third People’s Hospital, Nantong, Jiangsu, China), AuthorCompanyExt(id=1238891100761543133, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, companyId=1238891100740571608, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=南通市第三人民医院，南通大学附属南通第三医院检验科，江苏 南通)])]), Author(id=1238891103559144019, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, orderNo=3, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=0, email=null, emailSecond=null, emailThird=null, correspondingAuthor=0, authorType=1, ext={EN=AuthorExt(id=1238891103726916195, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, authorId=1238891103559144019, language=EN, stringName=Yiquan ZHANG, firstName=Yiquan, middleName=null, lastName=ZHANG, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=null, address=Department of Clinical Laboratory, Affiliated Nantong Hospital 3 of Nantong University, Nantong Third People’s Hospital, Nantong, Jiangsu, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1238891103852745328, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, authorId=1238891103559144019, language=CN, stringName=张义全, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=null, address=南通市第三人民医院，南通大学附属南通第三医院检验科，江苏 南通, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1238891100740571608, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, xref=null, ext=[AuthorCompanyExt(id=1238891100753154524, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, companyId=1238891100740571608, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Department of Clinical Laboratory, Affiliated Nantong Hospital 3 of Nantong University, Nantong Third People’s Hospital, Nantong, Jiangsu, China), AuthorCompanyExt(id=1238891100761543133, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, companyId=1238891100740571608, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=南通市第三人民医院，南通大学附属南通第三医院检验科，江苏 南通)])]), Author(id=1238891103991157370, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, orderNo=4, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=0, email=zm2860015024@163.com, emailSecond=null, emailThird=null, correspondingAuthor=1, authorType=1, ext={EN=AuthorExt(id=1238891104116986502, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, authorId=1238891103991157370, language=EN, stringName=Miaomiao ZHANG, firstName=Miaomiao, middleName=null, lastName=ZHANG, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=*, address=Department of Clinical Laboratory, Affiliated Nantong Hospital 3 of Nantong University, Nantong Third People’s Hospital, Nantong, Jiangsu, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1238891104230232724, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, authorId=1238891103991157370, language=CN, stringName=张苗苗, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=*, address=南通市第三人民医院，南通大学附属南通第三医院检验科，江苏 南通, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1238891100740571608, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, xref=null, ext=[AuthorCompanyExt(id=1238891100753154524, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, companyId=1238891100740571608, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Department of Clinical Laboratory, Affiliated Nantong Hospital 3 of Nantong University, Nantong Third People’s Hospital, Nantong, Jiangsu, China), AuthorCompanyExt(id=1238891100761543133, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, companyId=1238891100740571608, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=南通市第三人民医院，南通大学附属南通第三医院检验科，江苏 南通)])]), Author(id=1238891104356061856, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, orderNo=5, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=0, email=rainman78@163.com, emailSecond=null, emailThird=null, correspondingAuthor=1, authorType=1, ext={EN=AuthorExt(id=1238891104490279601, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, authorId=1238891104356061856, language=EN, stringName=Renfei LU, firstName=Renfei, middleName=null, lastName=LU, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=*, address=Department of Clinical Laboratory, Affiliated Nantong Hospital 3 of Nantong University, Nantong Third People’s Hospital, Nantong, Jiangsu, China, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=1238891104616108734, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, authorId=1238891104356061856, language=CN, stringName=陆仁飞, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=*, address=南通市第三人民医院，南通大学附属南通第三医院检验科，江苏 南通, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=[AuthorCompany(id=1238891100740571608, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, xref=null, ext=[AuthorCompanyExt(id=1238891100753154524, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, companyId=1238891100740571608, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Department of Clinical Laboratory, Affiliated Nantong Hospital 3 of Nantong University, Nantong Third People’s Hospital, Nantong, Jiangsu, China), AuthorCompanyExt(id=1238891100761543133, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, companyId=1238891100740571608, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=南通市第三人民医院，南通大学附属南通第三医院检验科，江苏 南通)])])], keywords=[Keyword(id=1238891104771297995, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, language=EN, orderNo=1, keyword=Vibrio parahaemolyticus), Keyword(id=1238891104909710037, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, language=EN, orderNo=2, keyword=quorum sensing), Keyword(id=1238891105010373338, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, language=EN, orderNo=3, keyword=GepA), Keyword(id=1238891105081676514, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, language=EN, orderNo=4, keyword=transcriptional regulation), Keyword(id=1238891105165562604, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, language=CN, orderNo=1, keyword=副溶血弧菌), Keyword(id=1238891105312363255, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, language=CN, orderNo=2, keyword=群体感应系统), Keyword(id=1238891105433998076, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, language=CN, orderNo=3, keyword=GepA), Keyword(id=1238891105534661382, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, language=CN, orderNo=4, keyword=转录调控)], refs=[Reference(id=1238891108936242074, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2025, volume=41, issue=2, pageStart=45, pageEnd=null, url=null, language=null, rfNumber=[1], rfOrder=0, authorNames=ZAKARIA NH, RAHIM NDE ABD, ROSILAN NF, SUNG YY, WAIHO K, HARUN S, ZAINAL ABIDIN RA, AFIQAH-ALENG N, journalName=World Journal of Microbiology and Biotechnology, refType=null, unstructuredReference=ZAKARIA NH, RAHIM NDE ABD, ROSILAN NF, SUNG YY, WAIHO K, HARUN S, ZAINAL ABIDIN RA, AFIQAH-ALENG N. Global landscape of Vibrio parahaemolyticus research: a bibliometric analysis[J]. World Journal of Microbiology and Biotechnology, 2025, 41(2): 45., articleTitle=Global landscape of Vibrio parahaemolyticus research: a bibliometric analysis, refAbstract=null), Reference(id=1238891109045293985, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2010, volume=57, issue=3/4, pageStart=179, pageEnd=182, url=null, language=null, rfNumber=[2], rfOrder=1, authorNames=SHIMOHATA T, TAKAHASHI A, journalName=Journal of Medical Investigation, refType=null, unstructuredReference=SHIMOHATA T, TAKAHASHI A. Diarrhea induced by infection of Vibrio parahaemolyticus [J]. Journal of Medical Investigation, 2010, 57(3/4): 179-182., articleTitle=Diarrhea induced by infection of Vibrio parahaemolyticus, refAbstract=null), Reference(id=1238891109162734506, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2010, volume=78, issue=4, pageStart=1772, pageEnd=1780, url=null, language=null, rfNumber=[3], rfOrder=2, authorNames=HIYOSHI H, KODAMA T, IIDA T, HONDA T, journalName=Infection and Immunity, refType=null, unstructuredReference=HIYOSHI H, KODAMA T, IIDA T, HONDA T. Contribution of Vibrio parahaemolyticus virulence factors to cytotoxicity, enterotoxicity, and lethality in mice[J]. Infection and Immunity, 2010, 78(4): 1772-1780., articleTitle=Contribution of Vibrio parahaemolyticus virulence factors to cytotoxicity, enterotoxicity, and lethality in mice, refAbstract=null), Reference(id=1238891109242426286, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2024, volume=175, issue=null, pageStart=113671, pageEnd=null, url=null, language=null, rfNumber=[4], rfOrder=3, authorNames=ASHIKUR RAHMAN M, AKTER S, ASHRAFUDOULLA M, ANAMUL HASAN CHOWDHURY M, UDDIN MAHAMUD AGMS, PARK SH, HA SD, journalName=Food Research International, refType=null, unstructuredReference=ASHIKUR RAHMAN M, AKTER S, ASHRAFUDOULLA M, ANAMUL HASAN CHOWDHURY M, UDDIN MAHAMUD AGMS, PARK SH, HA SD. Insights into the mechanisms and key factors influencing biofilm formation by Aeromonas hydrophila in the food industry: a comprehensive review and bibliometric analysis[J]. Food Research International, 2024, 175: 113671., articleTitle=Insights into the mechanisms and key factors influencing biofilm formation by Aeromonas hydrophila in the food industry: a comprehensive review and bibliometric analysis, refAbstract=null), Reference(id=1238891109338895283, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2021, volume=61, issue=11, pageStart=1827, pageEnd=1851, url=null, language=null, rfNumber=[5], rfOrder=4, authorNames=ASHRAFUDOULLA M, MIZAN MFR, PARK SH, HA SD, journalName=Critical Reviews in Food Science and Nutrition, refType=null, unstructuredReference=ASHRAFUDOULLA M, MIZAN MFR, PARK SH, HA SD. Current and future perspectives for controlling Vibrio biofilms in the seafood industry: a comprehensive review[J]. Critical Reviews in Food Science and Nutrition, 2021, 61(11): 1827-1851., articleTitle=Current and future perspectives for controlling Vibrio biofilms in the seafood industry: a comprehensive review, refAbstract=null), Reference(id=1238891109460530104, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2017, volume=15, issue=5, pageStart=271, pageEnd=284, url=null, language=null, rfNumber=[6], rfOrder=5, authorNames=JENAL U, REINDERS A, LORI C, journalName=Nature Reviews Microbiology, refType=null, unstructuredReference=JENAL U, REINDERS A, LORI C. Cyclic di-GMP: second messenger extraordinaire[J]. Nature Reviews Microbiology, 2017, 15(5): 271-284., articleTitle=Cyclic di-GMP: second messenger extraordinaire, refAbstract=null), Reference(id=1238891109561193407, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2013, volume=42, issue=1, pageStart=305, pageEnd=341, url=null, language=null, rfNumber=[7], rfOrder=6, authorNames=KALIA D, MEREY G, NAKAYAMA S, ZHENG Y, ZHOU J, LUO YL, GUO M, ROEMBKE BT, SINTIM HO, journalName=Chemical Society Reviews, refType=null, unstructuredReference=KALIA D, MEREY G, NAKAYAMA S, ZHENG Y, ZHOU J, LUO YL, GUO M, ROEMBKE BT, SINTIM HO. Nucleotide, c-di-GMP, c-di-AMP, cGMP, cAMP, (p)ppGpp signaling in bacteria and implications in pathogenesis[J]. Chemical Society Reviews, 2013, 42(1): 305-341., articleTitle=Nucleotide, c-di-GMP, c-di-AMP, cGMP, cAMP, (p)ppGpp signaling in bacteria and implications in pathogenesis, refAbstract=null), Reference(id=1238891109678633926, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2022, volume=27, issue=3, pageStart=157, pageEnd=172, url=null, language=null, rfNumber=[8], rfOrder=7, authorNames=HOMMA M, KOJIMA S, journalName=Genes to Cells, refType=null, unstructuredReference=HOMMA M, KOJIMA S. Roles of the second messenger c-di-GMP in bacteria: focusing on the topics of flagellar regulation and Vibrio spp.[J]. Genes to Cells, 2022, 27(3): 157-172., articleTitle=Roles of the second messenger c-di-GMP in bacteria: focusing on the topics of flagellar regulation and Vibrio spp, refAbstract=null), Reference(id=1238891109787685838, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2011, volume=108, issue=44, pageStart=18079, pageEnd=18084, url=null, language=null, rfNumber=[9], rfOrder=8, authorNames=TRIMBLE MJ, McCARTER LL, journalName=Proceedings of the National Academy of Sciences of the United States of America, refType=null, unstructuredReference=TRIMBLE MJ, McCARTER LL. Bis-(3′-5′)-cyclic dimeric GMP-linked quorum sensing controls swarming in Vibrio parahaemolyticus [J]. Proceedings of the National Academy of Sciences of the United States of America, 2011, 108(44): 18079-18084., articleTitle=Bis-(3′-5′)-cyclic dimeric GMP-linked quorum sensing controls swarming in Vibrio parahaemolyticus, refAbstract=null), Reference(id=1238891109909320661, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2007, volume=189, issue=11, pageStart=4094, pageEnd=4107, url=null, language=null, rfNumber=[10], rfOrder=9, authorNames=KIM YK, McCARTER LL, journalName=Journal of Bacteriology, refType=null, unstructuredReference=KIM YK, McCARTER LL. ScrG, a GGDEF-EAL protein, participates in regulating swarming and sticking in Vibrio parahaemolyticus [J]. Journal of Bacteriology, 2007, 189(11): 4094-4107., articleTitle=ScrG, a GGDEF-EAL protein, participates in regulating swarming and sticking in Vibrio parahaemolyticus, refAbstract=null), Reference(id=1238891109997401051, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2021, volume=203, issue=13, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[11], rfOrder=10, authorNames=MARTÍNEZ-MÉNDEZ R, CAMACHO-HERNÁNDEZ DA, SULVARÁN-GUEL E, ZAMORANO-SÁNCHEZ D, journalName=Journal of Bacteriology, refType=null, unstructuredReference=MARTÍNEZ-MÉNDEZ R, CAMACHO-HERNÁNDEZ DA, SULVARÁN-GUEL E, ZAMORANO-SÁNCHEZ D. A trigger phosphodiesterase modulates the global c-di-GMP pool, motility, and biofilm formation in Vibrio parahaemolyticus [J]. Journal of Bacteriology, 2021, 203(13): e0004621., articleTitle=A trigger phosphodiesterase modulates the global c-di-GMP pool, motility, and biofilm formation in Vibrio parahaemolyticus, refAbstract=null), Reference(id=1238891110085481440, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2023, volume=42, issue=11, pageStart=113456, pageEnd=null, url=null, language=null, rfNumber=[12], rfOrder=11, authorNames=WU X, ZHOU LT, YE C, ZHA ZZ, LI CC, FENG C, ZHANG Y, JIN Q, PAN JY, journalName=Cell Reports, refType=null, unstructuredReference=WU X, ZHOU LT, YE C, ZHA ZZ, LI CC, FENG C, ZHANG Y, JIN Q, PAN JY. Destruction of self-derived PAMP via T3SS2 effector VopY to subvert PAMP-triggered immunity mediates Vibrio parahaemolyticus pathogenicity[J]. Cell Reports, 2023, 42(11): 113456., articleTitle=Destruction of self-derived PAMP via T3SS2 effector VopY to subvert PAMP-triggered immunity mediates Vibrio parahaemolyticus pathogenicity, refAbstract=null), Reference(id=1238891110257447910, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2022, volume=88, issue=6, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[13], rfOrder=12, authorNames=ZHONG XJ, LU Z, WANG F, YAO N, SHI MT, YANG MH, journalName=Applied and Environmental Microbiology, refType=null, unstructuredReference=ZHONG XJ, LU Z, WANG F, YAO N, SHI MT, YANG MH. Characterization of GefA, a GGEEF domain-containing protein that modulates Vibrio parahaemolyticus motility, biofilm formation, and virulence[J]. Applied and Environmental Microbiology, 2022, 88(6): e0223921., articleTitle=Characterization of GefA, a GGEEF domain-containing protein that modulates Vibrio parahaemolyticus motility, biofilm formation, and virulence, refAbstract=null), Reference(id=1238891110383277037, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2020, volume=202, issue=6, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[14], rfOrder=13, authorNames=KIMBROUGH JH, CRIBBS JT, McCARTER LL, journalName=Journal of Bacteriology, refType=null, unstructuredReference=KIMBROUGH JH, CRIBBS JT, McCARTER LL. Homologous c-di-GMP-binding scr transcription factors orchestrate biofilm development in Vibrio parahaemolyticus [J]. Journal of Bacteriology, 2020, 202(6): e00723-19., articleTitle=Homologous c-di-GMP-binding scr transcription factors orchestrate biofilm development in Vibrio parahaemolyticus, refAbstract=null), Reference(id=1238891110467163121, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2021, volume=203, issue=4, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[15], rfOrder=14, authorNames=KIMBROUGH JH, McCARTER LL, journalName=Journal of Bacteriology, refType=null, unstructuredReference=KIMBROUGH JH, McCARTER LL. Identification of three new GGDEF and EAL domain-containing proteins participating in the scr surface colonization regulatory network in Vibrio parahaemolyticus [J]. Journal of Bacteriology, 2021, 203(4): e00409-20., articleTitle=Identification of three new GGDEF and EAL domain-containing proteins participating in the scr surface colonization regulatory network in Vibrio parahaemolyticus, refAbstract=null), Reference(id=1238891110601380859, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2025, volume=933, issue=null, pageStart=148968, pageEnd=null, url=null, language=null, rfNumber=[16], rfOrder=15, authorNames=ZHOU YN, CHANG JY, ZHANG MM, LI X, LUO X, LI WP, TIAN ZK, ZHANG N, NI B, ZHANG YQ, LU RF, journalName=Gene, refType=null, unstructuredReference=ZHOU YN, CHANG JY, ZHANG MM, LI X, LUO X, LI WP, TIAN ZK, ZHANG N, NI B, ZHANG YQ, LU RF. GefB, a GGDEF domain-containing protein, affects motility and biofilm formation of Vibrio parahaemolyticus and is regulated by quorum sensing regulators[J]. Gene, 2025, 933: 148968., articleTitle=GefB, a GGDEF domain-containing protein, affects motility and biofilm formation of Vibrio parahaemolyticus and is regulated by quorum sensing regulators, refAbstract=null), Reference(id=1238891110739792893, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2024, volume=195, issue=null, pageStart=106882, pageEnd=null, url=null, language=null, rfNumber=[17], rfOrder=16, authorNames=ZHOU YN, CHANG JY, ZHANG MM, LI X, YANG WH, HU LF, ZHOU DS, NI B, LU RF, ZHANG YQ, journalName=Microbial Pathogenesis, refType=null, unstructuredReference=ZHOU YN, CHANG JY, ZHANG MM, LI X, YANG WH, HU LF, ZHOU DS, NI B, LU RF, ZHANG YQ. VPA0198, a GGDEF domain-containing protein, affects the motility and biofilm formation of Vibrio parahaemolyticus and is regulated by quorum sensing associated regulators[J]. Microbial Pathogenesis, 2024, 195: 106882., articleTitle=VPA0198, a GGDEF domain-containing protein, affects the motility and biofilm formation of Vibrio parahaemolyticus and is regulated by quorum sensing associated regulators, refAbstract=null), Reference(id=1238891110907564041, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2011, volume=193, issue=16, pageStart=4224, pageEnd=4237, url=null, language=null, rfNumber=[18], rfOrder=17, authorNames=GODE-POTRATZ CJ, McCARTER LL, journalName=Journal of Bacteriology, refType=null, unstructuredReference=GODE-POTRATZ CJ, McCARTER LL. Quorum sensing and silencing in Vibrio parahaemolyticus [J]. Journal of Bacteriology, 2011, 193(16): 4224-4237., articleTitle=Quorum sensing and silencing in Vibrio parahaemolyticus, refAbstract=null), Reference(id=1238891112404930580, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2023, volume=851, issue=null, pageStart=146980, pageEnd=null, url=null, language=null, rfNumber=[19], rfOrder=18, authorNames=CHEN L, ZHANG MM, LI X, WU QM, XUE XF, ZHANG TT, LU RF, ZHANG YQ, journalName=Gene, refType=null, unstructuredReference=CHEN L, ZHANG MM, LI X, WU QM, XUE XF, ZHANG TT, LU RF, ZHANG YQ. AphA directly activates the transcription of polysaccharide biosynthesis gene scvE in Vibrio parahaemolyticus [J]. Gene, 2023, 851: 146980., articleTitle=AphA directly activates the transcription of polysaccharide biosynthesis gene scvE in Vibrio parahaemolyticus, refAbstract=null), Reference(id=1238891112547536919, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2019, volume=21, issue=3, pageStart=1054, pageEnd=1067, url=null, language=null, rfNumber=[20], rfOrder=19, authorNames=ZHANG YQ, HU LH, QIU Y, OSEI-ADJEI G, TANG H, ZHANG Y, ZHANG R, SHENG XM, XU SG, YANG WH, YANG HY, YIN Z, YANG RF, HUANG XX, ZHOU DS, journalName=Environmental Microbiology, refType=null, unstructuredReference=ZHANG YQ, HU LH, QIU Y, OSEI-ADJEI G, TANG H, ZHANG Y, ZHANG R, SHENG XM, XU SG, YANG WH, YANG HY, YIN Z, YANG RF, HUANG XX, ZHOU DS. QsvR integrates into quorum sensing circuit to control Vibrio parahaemolyticus virulence[J]. Environmental Microbiology, 2019, 21(3): 1054-1067., articleTitle=QsvR integrates into quorum sensing circuit to control Vibrio parahaemolyticus virulence, refAbstract=null), Reference(id=1238891112673366044, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2023, volume=14, issue=null, pageStart=1123524, pageEnd=null, url=null, language=null, rfNumber=[21], rfOrder=20, authorNames=ZHANG YQ, XUE XF, SUN FJ, LI X, ZHANG MM, WU QM, ZHANG TT, LUO X, LU RF, journalName=Frontiers in Microbiology, refType=null, unstructuredReference=ZHANG YQ, XUE XF, SUN FJ, LI X, ZHANG MM, WU QM, ZHANG TT, LUO X, LU RF. Quorum sensing and QsvR tightly control the transcription of vpa0607 encoding an active RNase II-type protein in Vibrio parahaemolyticus [J]. Frontiers in Microbiology, 2023, 14: 1123524., articleTitle=Quorum sensing and QsvR tightly control the transcription of vpa0607 encoding an active RNase II-type protein in Vibrio parahaemolyticus, refAbstract=null), Reference(id=1238891112753057826, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2023, volume=14, issue=null, pageStart=1079653, pageEnd=null, url=null, language=null, rfNumber=[22], rfOrder=21, authorNames=ZHANG MM, XUE XF, LI X, WU QM, ZHANG TT, YANG WH, HU LF, ZHOU DS, LU RF, ZHANG YQ, journalName=Frontiers in Microbiology, refType=null, unstructuredReference=ZHANG MM, XUE XF, LI X, WU QM, ZHANG TT, YANG WH, HU LF, ZHOU DS, LU RF, ZHANG YQ. QsvR and OpaR coordinately repress biofilm formation by Vibrio parahaemolyticus [J]. Frontiers in Microbiology, 2023, 14: 1079653., articleTitle=QsvR and OpaR coordinately repress biofilm formation by Vibrio parahaemolyticus, refAbstract=null), Reference(id=1238891112874692648, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2023, volume=174, issue=null, pageStart=105947, pageEnd=null, url=null, language=null, rfNumber=[23], rfOrder=22, authorNames=ZHANG MM, XUE XF, LI X, LUO X, WU QM, ZHANG TT, YANG WH, HU LF, ZHOU DS, LU RF, ZHANG YQ, journalName=Microbial Pathogenesis, refType=null, unstructuredReference=ZHANG MM, XUE XF, LI X, LUO X, WU QM, ZHANG TT, YANG WH, HU LF, ZHOU DS, LU RF, ZHANG YQ. QsvR represses the transcription of polar flagellum genes in Vibrio parahaemolyticus [J]. Microbial Pathogenesis, 2023, 174: 105947., articleTitle=QsvR represses the transcription of polar flagellum genes in Vibrio parahaemolyticus, refAbstract=null), Reference(id=1238891112983744561, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2018, volume=8, issue=null, pageStart=291, pageEnd=null, url=null, language=null, rfNumber=[24], rfOrder=23, authorNames=ZHANG YQ, HU LF, OSEI-ADJEI G, ZHANG Y, YANG WH, YIN Z, LU RY, SHENG XM, YANG RF, HUANG XX, ZHOU DS, journalName=Frontiers in Cellular and Infection Microbiology, refType=null, unstructuredReference=ZHANG YQ, HU LF, OSEI-ADJEI G, ZHANG Y, YANG WH, YIN Z, LU RY, SHENG XM, YANG RF, HUANG XX, ZHOU DS. Autoregulation of ToxR and its regulatory actions on major virulence gene loci in Vibrio parahaemolyticus [J]. Frontiers in Cellular and Infection Microbiology, 2018, 8: 291., articleTitle=Autoregulation of ToxR and its regulatory actions on major virulence gene loci in Vibrio parahaemolyticus, refAbstract=null), Reference(id=1238891113101185079, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2017, volume=8, issue=null, pageStart=2005, pageEnd=null, url=null, language=null, rfNumber=[25], rfOrder=24, authorNames=ZHANG YQ, GAO H, OSEI-ADJEI G, ZHANG Y, YANG WH, YANG HY, YIN Z, HUANG XX, ZHOU DS, journalName=Frontiers in Microbiology, refType=null, unstructuredReference=ZHANG YQ, GAO H, OSEI-ADJEI G, ZHANG Y, YANG WH, YANG HY, YIN Z, HUANG XX, ZHOU DS. Transcriptional regulation of the type VI secretion system 1 genes by quorum sensing and ToxR in Vibrio parahaemolyticus [J]. Frontiers in Microbiology, 2017, 8: 2005., articleTitle=Transcriptional regulation of the type VI secretion system 1 genes by quorum sensing and ToxR in Vibrio parahaemolyticus, refAbstract=null), Reference(id=1238891113247985728, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2017, volume=8, issue=39, pageStart=65809, pageEnd=65822, url=null, language=null, rfNumber=[26], rfOrder=25, authorNames=OSEI-ADJEI G, GAO H, ZHANG Y, ZHANG LY, YANG WH, YANG HY, YIN Z, HUANG XX, ZHANG YQ, ZHOU DS, journalName=Oncotarget, refType=null, unstructuredReference=OSEI-ADJEI G, GAO H, ZHANG Y, ZHANG LY, YANG WH, YANG HY, YIN Z, HUANG XX, ZHANG YQ, ZHOU DS. Regulatory actions of ToxR and CalR on their own genes and type III secretion system 1 in Vibrio parahaemolyticus [J]. Oncotarget, 2017, 8(39): 65809-65822., articleTitle=Regulatory actions of ToxR and CalR on their own genes and type III secretion system 1 in Vibrio parahaemolyticus, refAbstract=null), Reference(id=1238891113378009157, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2014, volume=54, issue=5, pageStart=525, pageEnd=531, url=null, language=null, rfNumber=[27], rfOrder=26, authorNames=黄倩, 张义全, 胡小许, 王丽, 杨瑞馥, 钟青萍, 周冬生, 黎晓敏, journalName=微生物学报, refType=null, unstructuredReference=黄倩, 张义全, 胡小许, 王丽, 杨瑞馥, 钟青萍, 周冬生, 黎晓敏. AphA蛋白促进副溶血弧菌c-di-GMP合成和生物膜形成[J]. 微生物学报, 2014, 54(5): 525-531., articleTitle=AphA蛋白促进副溶血弧菌c-di-GMP合成和生物膜形成, refAbstract=null), Reference(id=1238891113436729419, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2014, volume=54, issue=5, pageStart=525, pageEnd=531, url=null, language=null, rfNumber=[27], rfOrder=27, authorNames=HUANG Q, ZHANG YQ, HU XX, WANG L, YANG RF, ZHONG QP, ZHOU DS, LI XM, journalName=Acta Microbiologica Sinica, refType=null, unstructuredReference=HUANG Q, ZHANG YQ, HU XX, WANG L, YANG RF, ZHONG QP, ZHOU DS, LI XM. AphA is an activator of c-di-GMP synthesis and biofilm formation in Vibrio parahaemolyticus [J]. Acta Microbiologica Sinica, 2014, 54(5): 525-531 (in Chinese)., articleTitle=null, refAbstract=null), Reference(id=1238891113566752850, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2022, volume=38, issue=12, pageStart=4719, pageEnd=4730, url=null, language=null, rfNumber=[28], rfOrder=28, authorNames=张苗苗, 薛星帆, 孙君芳, 吴齐敏, 李雪, 周冬生, 倪斌, 陆仁飞, 张义全, journalName=生物工程学报, refType=null, unstructuredReference=张苗苗, 薛星帆, 孙君芳, 吴齐敏, 李雪, 周冬生, 倪斌, 陆仁飞, 张义全. ToxR负调控副溶血弧菌中c-di-GMP的合成[J]. 生物工程学报, 2022, 38(12): 4719-4730., articleTitle=ToxR负调控副溶血弧菌中c-di-GMP的合成, refAbstract=null), Reference(id=1238891113667416152, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2022, volume=38, issue=12, pageStart=4719, pageEnd=4730, url=null, language=null, rfNumber=[28], rfOrder=29, authorNames=ZHANG MM, XUE XF, SUN JF, WU QM, LI X, ZHOU DS, NI B, LU RF, ZHANG YQ, journalName=Chinese Journal of Biotechnology, refType=null, unstructuredReference=ZHANG MM, XUE XF, SUN JF, WU QM, LI X, ZHOU DS, NI B, LU RF, ZHANG YQ. ToxR represses the synthesis of c-di-GMP in Vibrio parahaemolyticus [J]. Chinese Journal of Biotechnology, 2022, 38(12): 4719-4730 (in Chinese)., articleTitle=null, refAbstract=null), Reference(id=1238891113768079451, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2025, volume=207, issue=5, pageStart=99, pageEnd=null, url=null, language=null, rfNumber=[29], rfOrder=30, authorNames=ZHANG MM, ZHU YR, LI X, LUO X, SUN H, XIONG SH, LU RF, ZHANG YQ, journalName=Archives of Microbiology, refType=null, unstructuredReference=ZHANG MM, ZHU YR, LI X, LUO X, SUN H, XIONG SH, LU RF, ZHANG YQ. GepA, a GGDEF-EAL protein, regulates biofilm formation and swimming motility in Vibrio parahaemolyticus [J]. Archives of Microbiology, 2025, 207(5): 99., articleTitle=GepA, a GGDEF-EAL protein, regulates biofilm formation and swimming motility in Vibrio parahaemolyticus, refAbstract=null), Reference(id=1238891113919074402, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2021, volume=12, issue=null, pageStart=676436, pageEnd=null, url=null, language=null, rfNumber=[30], rfOrder=31, authorNames=ZHANG YQ, QIU Y, GAO H, SUN JF, LI X, ZHANG MM, XUE XF, YANG WH, NI B, HU LF, YIN Z, LU RF, ZHOU DS, journalName=Frontiers in Microbiology, refType=null, unstructuredReference=ZHANG YQ, QIU Y, GAO H, SUN JF, LI X, ZHANG MM, XUE XF, YANG WH, NI B, HU LF, YIN Z, LU RF, ZHOU DS. OpaR controls the metabolism of c-di-GMP in Vibrio parahaemolyticus [J]. Frontiers in Microbiology, 2021, 12: 676436., articleTitle=OpaR controls the metabolism of c-di-GMP in Vibrio parahaemolyticus, refAbstract=null), Reference(id=1238891114028126308, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2015, volume=10, issue=12, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=[31], rfOrder=32, authorNames=MA YJ, SUN XH, XU XY, ZHAO Y, PAN YJ, HWANG CA, WU VCH, journalName=PLoS One, refType=null, unstructuredReference=MA YJ, SUN XH, XU XY, ZHAO Y, PAN YJ, HWANG CA, WU VCH. Investigation of reference genes in Vibrio parahaemolyticus for gene expression analysis using quantitative RT-PCR[J]. PLoS One, 2015, 10(12): e0144362., articleTitle=Investigation of reference genes in Vibrio parahaemolyticus for gene expression analysis using quantitative RT-PCR, refAbstract=null), Reference(id=1238891114166538346, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2021, volume=13, issue=1, pageStart=15, pageEnd=null, url=null, language=null, rfNumber=[32], rfOrder=33, authorNames=ZHANG YQ, QIU Y, XUE XF, ZHANG MM, SUN JF, LI X, HU LF, YIN Z, YANG WH, LU RF, ZHOU DS, journalName=Gut Pathogens, refType=null, unstructuredReference=ZHANG YQ, QIU Y, XUE XF, ZHANG MM, SUN JF, LI X, HU LF, YIN Z, YANG WH, LU RF, ZHOU DS. Transcriptional regulation of the virulence genes and the biofilm formation associated operons in Vibrio parahaemolyticus [J]. Gut Pathogens, 2021, 13(1): 15., articleTitle=Transcriptional regulation of the virulence genes and the biofilm formation associated operons in Vibrio parahaemolyticus, refAbstract=null), Reference(id=1238891114283978866, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2022, volume=35, issue=9, pageStart=821, pageEnd=829, url=null, language=null, rfNumber=[33], rfOrder=34, authorNames=XUE XF, ZHNAG MM, SUN JF, LI X, WU QM, YIN Z, YANG WH, NI B, HU LF, ZHOU DS, LU RF, ZHANG YQ, journalName=Biomedical and Environmental Sciences, refType=null, unstructuredReference=XUE XF, ZHNAG MM, SUN JF, LI X, WU QM, YIN Z, YANG WH, NI B, HU LF, ZHOU DS, LU RF, ZHANG YQ. H-NS represses biofilm formation and c-di-GMP synthesis in Vibrio parahaemolyticus [J]. Biomedical and Environmental Sciences, 2022, 35(9): 821-829., articleTitle=H-NS represses biofilm formation and c-di-GMP synthesis in Vibrio parahaemolyticus, refAbstract=null), Reference(id=1238891114414002298, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2021, volume=79, issue=1, pageStart=25, pageEnd=null, url=null, language=null, rfNumber=[34], rfOrder=35, authorNames=LI X, SUN JF, ZHANG MM, XUE XF, WU QM, YANG WH, YIN Z, ZHOU DS, LU RF, ZHANG YQ, journalName=Current Microbiology, refType=null, unstructuredReference=LI X, SUN JF, ZHANG MM, XUE XF, WU QM, YANG WH, YIN Z, ZHOU DS, LU RF, ZHANG YQ. The effect of salinity on biofilm formation and c-di-GMP production in Vibrio parahaemolyticus [J]. Current Microbiology, 2021, 79(1): 25., articleTitle=The effect of salinity on biofilm formation and c-di-GMP production in Vibrio parahaemolyticus, refAbstract=null), Reference(id=1238891114560802948, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, doi=null, pmid=null, pmcid=null, year=2024, volume=3, issue=1, pageStart=42, pageEnd=56, url=null, language=null, rfNumber=[35], rfOrder=36, authorNames=LIU C, SHI R, JENSEN MS, ZHU JR, LIU JW, LIU XB, SUN D, LIU WJ, journalName=mLife, refType=null, unstructuredReference=LIU C, SHI R, JENSEN MS, ZHU JR, LIU JW, LIU XB, SUN D, LIU WJ. The global regulation of c-di-GMP and cAMP in bacteria[J]. mLife, 2024, 3(1): 42-56., articleTitle=The global regulation of c-di-GMP and cAMP in bacteria, refAbstract=null)], funds=[Fund(id=1238891108592309123, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, awardId=JCZ2025004, language=EN, fundingSource=Natural Science Foundation of Nantong(JCZ2025004), fundOrder=null, country=null), Fund(id=1238891108684583818, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, awardId=JCZ2025004, language=CN, fundingSource=南通市自然科学基金(JCZ2025004), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1238891100740571608, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, xref=null, ext=[AuthorCompanyExt(id=1238891100753154524, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, companyId=1238891100740571608, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Department of Clinical Laboratory, Affiliated Nantong Hospital 3 of Nantong University, Nantong Third People’s Hospital, Nantong, Jiangsu, China), AuthorCompanyExt(id=1238891100761543133, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, companyId=1238891100740571608, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=南通市第三人民医院，南通大学附属南通第三医院检验科，江苏 南通)])], figs=[ArticleFig(id=1238891105790513949, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, language=EN, label=Figure 1, caption=Positive regulation of gepA by AphA. The negative and positive numbers represent the nucleotide positions upstream and downstream of the first base of the start codon, respectively. A: qPCR (The relative mRNA level of gepA was compared between ΔaphA and WT); B: Luminescence assay [The regulatory DNA region of gepA was cloned into the pBBRlux vector, and then introduced into ΔaphA and WT, to test the luminescence activity of each strain. The luminescence activity (RLU) was calculated as light units/OD600]; C: Primer extension (Lanes C, T, A, and G represent the Sanger sequencing reactions. The transcriptional start sites were indicated by arrows with nucleotides and positions); D: EMSA [The gepA DNA fragments were radioactively labeled with [γ-32P]-ATP (5 000 Ci/mmol), and then incubated with increasing amounts of His-AphA, followed by 4% (W/V) polyacrylamide gel electrophoresis. Results were analyzed by autoradiography after exposure to Fuji Medical X-ray film. Lanes 1, 2, 3, 4, 5, 6 and 7 contain 0, 19.5, 29.3, 39.0, 39.0, 39.0 and 0 pmol of His-AphA, respectively, and then lane 5 contains 2 pmol cold probe, lane 6 contains 2 pmol negative probe, and lane 7 contains 2 pmol unrelated protein]., figureFileSmall=dGQWgQeqCGr496Tg2eEFIg==, figureFileBig=55no01/5PxiKlYhGnDpl3Q==, tableContent=null), ArticleFig(id=1238891105928925988, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, language=CN, label=图1, caption=AphA正调控 gepA 的转录, figureFileSmall=dGQWgQeqCGr496Tg2eEFIg==, figureFileBig=55no01/5PxiKlYhGnDpl3Q==, tableContent=null), ArticleFig(id=1238891106054755121, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, language=EN, label=Figure 2, caption=Positive regulation of gepA by ToxR. The negative and positive numbers represent the nucleotide positions upstream and downstream of the first base of the start codon, respectively. A: qPCR; B: Luminescence assay; C: EMSA (lanes 1, 2, 3, 4, 5, 6 and 7 contain 0, 0.15, 0.45, 0.60, 0.60, 0.60 and 0 pmol of His-ToxR, respectively, and then lane 5 contains 2 pmol cold probe, lane 6 contains 2 pmol negative probe, and lane 7 contains 2 pmol unrelated protein)., figureFileSmall=2VSTuIkwRjBIqSIzEImt3A==, figureFileBig=sIZ8g4KPmK0Z/RvJK3+cNQ==, tableContent=null), ArticleFig(id=1238891106159612726, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, language=CN, label=图2, caption=ToxR正调控 gepA 的转录, figureFileSmall=2VSTuIkwRjBIqSIzEImt3A==, figureFileBig=sIZ8g4KPmK0Z/RvJK3+cNQ==, tableContent=null), ArticleFig(id=1238891106285441855, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, language=EN, label=Figure 3, caption=Negative regulation of gepA by QsvR. The negative and positive numbers represent the nucleotide positions upstream and downstream of the first base of the start codon, respectively. A: qPCR; B: Primer extension; C: EMSA [The regulatory DNA region of gepA was incubated with increasing amounts of purified His-QsvR, and then subjected to 6% (W/V) polyacrylamide gel electrophoresis. The DNA bands were visualized by the EB staining. Lanes 1, 2, 3, 4, 5, 6 and 7 contain 0, 0.021, 0.042, 0.063, 0.083, 0.110 and 0.130 pmol of His-QsvR, respectively]; D: Two-plasmid reporter assay [The pBAD33-qsvR plasmid or the empty pBAD33 vector and a recombinant lacZ plasmid were simultaneously introduced into the E. coli 100 λpir (Epicentre), and then the promoter activities (represented by Miller units) of each target gene in the cellular extracts were determined by a β-galactosidase Enzyme Assay System according to the manufacturer’s instructions]., figureFileSmall=+wfdvb/zhA99b8sCG9fXMw==, figureFileBig=67e40cYSbwGCfgRDnZ8sCw==, tableContent=null), ArticleFig(id=1238891106423853894, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, language=CN, label=图3, caption=QsvR负调控 gepA 的转录, figureFileSmall=+wfdvb/zhA99b8sCG9fXMw==, figureFileBig=67e40cYSbwGCfgRDnZ8sCw==, tableContent=null), ArticleFig(id=1238891107896054606, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, language=EN, label=Figure 4, caption=Cell density-dependent transcription of gepA. The WT strain was grown in HI broth at 37 ℃, and then the luminescence activity of bacterial cells was tested at different OD600 values., figureFileSmall=BMIzwm5/1aQ4mK+j2Wk34Q==, figureFileBig=SeAK5gbIao2JmyebPzm71g==, tableContent=null), ArticleFig(id=1238891108026078041, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, language=CN, label=图4, caption=gepA 呈密度依赖性转录, figureFileSmall=BMIzwm5/1aQ4mK+j2Wk34Q==, figureFileBig=SeAK5gbIao2JmyebPzm71g==, tableContent=null), ArticleFig(id=1238891108126741346, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, language=EN, label=Figure 5, caption=The transcriptional regulatory model of gepA regulated by QS-related regulatory elements., figureFileSmall=7Jyd67ZO9Sgc62arJ1UAlA==, figureFileBig=R5NEbLOiW8ps1EcSiaELDQ==, tableContent=null), ArticleFig(id=1238891108227404650, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, language=CN, label=图5, caption=QS相关调控子对 gepA 的转录调控模式图, figureFileSmall=7Jyd67ZO9Sgc62arJ1UAlA==, figureFileBig=R5NEbLOiW8ps1EcSiaELDQ==, tableContent=null), ArticleFig(id=1238891108323873650, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, language=EN, label=Table 1, caption=

Oligonucleotide primers used in this study

, figureFileSmall=null, figureFileBig=null, tableContent=
TargetsPrimer sequences (5′→3′)
qPCR
gepAGACCACCTCAATAGTTATCTG/TAAGTAGGCTTGGACATCTC
16S rRNAGACACGGTCCAGACTCCTAC/GGTGCTTCTTCTGTCGCTAAC
Lux
gepAGCGCGAGCTCCTCACACAACACTTTCTG/GCGCGGATCCAGACAATCACACCGATAG
Primer extension
gepACACACTAAAGGTCACAAGCAAG/AGACAATCACACCGATAG
LacZ fusion
gepAGCGCTCTAGACTCACACAACACTTTCTG/GCGCGAATTCAGACAATCACACCGATAG
EMSA
gepAGCGCTCTAGACTCACACAACACTTTCTG/GCGCGAATTCAGACAATCACACCGATAG
vp1687GCGCGTCGACGCATTATTGACGCCAGTATCG /GCGCTCTAGAGGCAACGGTGAGCAAAATC
), ArticleFig(id=1238891108399371128, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1238813313346359685, language=CN, label=表1, caption=

本研究所用引物

, figureFileSmall=null, figureFileBig=null, tableContent=
TargetsPrimer sequences (5′→3′)
qPCR
gepAGACCACCTCAATAGTTATCTG/TAAGTAGGCTTGGACATCTC
16S rRNAGACACGGTCCAGACTCCTAC/GGTGCTTCTTCTGTCGCTAAC
Lux
gepAGCGCGAGCTCCTCACACAACACTTTCTG/GCGCGGATCCAGACAATCACACCGATAG
Primer extension
gepACACACTAAAGGTCACAAGCAAG/AGACAATCACACCGATAG
LacZ fusion
gepAGCGCTCTAGACTCACACAACACTTTCTG/GCGCGAATTCAGACAATCACACCGATAG
EMSA
gepAGCGCTCTAGACTCACACAACACTTTCTG/GCGCGAATTCAGACAATCACACCGATAG
vp1687GCGCGTCGACGCATTATTGACGCCAGTATCG /GCGCTCTAGAGGCAACGGTGAGCAAAATC
)], attaches=null, journal=Journal(id=1192105720683257860, delFlag=0, nameCn=微生物学报, nameEn=Acta Microbiologica Sinica, nameHistory1=null, nameHistory2=null, issn=0001-6209, eissn=null, cn=11-1995/Q, coden=null, periodic=0, language=CN, oaType=null, ccby=null, superviseOffice=null, ownerOffice=null, pubOffice=null, editorOffice=null, officeType=null, aims=null, clcCode=null, officeProv=null, officeCity=null, officeAddr=null, officeZip=null, officeEmail=null, officePhone=null, editDirector=null, officeDirector=null, officeDirectorPhone=null, officeStaffNum=null, officeEmpNum=null, coverPicUrl=tNA7JigLZj/rxynSmzKgDQ==, journalPrice=null, startedYear=null, abbrevIsoEn=null, journalRemark=null, publicationField=null, createdTime=1762149752067, updatedTime=1762150746905, createdBy=18614031015, updatedBy=13701087609, firstLetterCn=A, firstLetterEn=A, subjectCode=Life Sciences, subjectName=Life Sciences, subjectCodeEn=Life Sciences, subjectNameEn=null, picCn=tNA7JigLZj/rxynSmzKgDQ==, picEn=R/d5eSUu8/o5mAGWCF3M5Q==, jcr=null, cjcr=null, exts=[JournalExt(id=1192109893441171829, language=CN, name=微生物学报, nameHistory1=null, nameHistory2=null, managedBy=, sponsoredBy=, publishedBy=, editorOffice=, officeProv=null, officeCity=null, officeAddr=, officeZip=, editDirector=, officeDirector=null, officePhone=null, coverPicUrl=null, journalRemark=, submitArticleUrl=null, websiteUrl=, createdTime=1762150746928, updatedTime=1762150746928, createdBy=13701087609, updatedBy=13701087609, submissionGuidelinesUrl=, submissionAuthorUrl=https://actamicro.ijournals.cn/actamicrocn/author/login, submissionEditorUrl=https://actamicro.ijournals.cn/actamicrocn/editor/login, submissionReviewUrl=https://actamicro.ijournals.cn/actamicrocn/reviewer/login, submissionCeEditorUrl=, submissionAeEditorUrl=, option={"copyright":""}), JournalExt(id=1192109893512474998, language=EN, name=Acta Microbiologica Sinica, nameHistory1=null, nameHistory2=null, managedBy=, sponsoredBy=, publishedBy=, editorOffice=, officeProv=null, officeCity=null, officeAddr=, officeZip=, editDirector=, officeDirector=null, officePhone=null, coverPicUrl=null, journalRemark=, submitArticleUrl=null, websiteUrl=, createdTime=1762150746944, updatedTime=1762150746944, createdBy=13701087609, updatedBy=13701087609, submissionGuidelinesUrl=, submissionAuthorUrl=https://actamicro.ijournals.cn/actamicrocn/author/login, submissionEditorUrl=https://actamicro.ijournals.cn/actamicrocn/editor/login, submissionReviewUrl=https://actamicro.ijournals.cn/actamicrocn/reviewer/login, submissionCeEditorUrl=, submissionAeEditorUrl=, option={"copyright":""})], databaseList=null, tenantJournalId=1192105938417971205, websiteList=[Website(id=1192106105867223981, webName=null, webTitle=null, webDomain=null, webCopyrigh=null, webIpcNo=null, seoTitle=null, seoKeywords=null, seoDescription=null, tenantJournalId=null, journalId=1192105938417971205, journalNameCn=null, journalNameEn=null, grayFlag=null, tenantId=1146029695717560320, platformId=null, journalGroupId=null, journalGroupNameCn=null, journalGroupNameEn=null, type=1, domain=https://castjournals.cast.org.cn/joweb/wswxb/CN, language=CN, createTime=1762149843899, createBy=18614031015, updateTime=1762149888800, updateBy=18614031015, name=微生物学报-中文, tplId=1146099689490845704, title=微生物学报, delFlag=0, indexPage=/home, props=[WebsiteProps(id=1192107120863626198, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1192106105867223981, code=articleTextType, value=kx, createTime=1762150085893, updateTime=1762150085893, creator=18614031015, updator=18614031015), WebsiteProps(id=1192107120834266067, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1192106105867223981, code=banner, value=null, createTime=1762150085886, updateTime=1762150085886, creator=18614031015, updator=18614031015), WebsiteProps(id=1192107120892986329, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1192106105867223981, code=grayFlag, value=0, createTime=1762150085900, updateTime=1762150085900, creator=18614031015, updator=18614031015), WebsiteProps(id=1192107120825877458, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1192106105867223981, code=logo, value=https://castjournals.cast.org.cn/joweb/wswxb/CN/file/pic?fileId=FOz4Ks7dC79FYnCEBIlMdw==, createTime=1762150085884, updateTime=1762150085884, creator=18614031015, updator=18614031015), WebsiteProps(id=1192107120905569243, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1192106105867223981, code=minRunFlag, value=0, createTime=1762150085903, updateTime=1762150085903, creator=18614031015, updator=18614031015), WebsiteProps(id=1192107120846848981, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1192106105867223981, code=picServerUrl, value=https://castjournals.cast.org.cn/joweb/wswxb/CN/file/pic, createTime=1762150085889, updateTime=1762150085889, creator=18614031015, updator=18614031015), WebsiteProps(id=1192107120897180634, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1192106105867223981, code=silenceFlag, value=0, createTime=1762150085901, updateTime=1762150085901, creator=18614031015, updator=18614031015), WebsiteProps(id=1192107120842654676, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1192106105867223981, code=staticResourcePath, value=https://castjournals.cast.org.cn/joweb/cast_kjdb_cn_619/, createTime=1762150085888, updateTime=1762150085888, creator=18614031015, updator=18614031015), WebsiteProps(id=1192107120872014807, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1192106105867223981, code=themeColor, value=null, createTime=1762150085895, updateTime=1762150085895, creator=18614031015, updator=18614031015), WebsiteProps(id=1192107120880403416, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1192106105867223981, code=themeStyle, value=null, createTime=1762150085897, updateTime=1762150085897, creator=18614031015, updator=18614031015)]), Website(id=1192106106018218929, webName=null, webTitle=null, webDomain=null, webCopyrigh=null, webIpcNo=null, seoTitle=null, seoKeywords=null, seoDescription=null, tenantJournalId=null, journalId=1192105938417971205, journalNameCn=null, journalNameEn=null, grayFlag=null, tenantId=1146029695717560320, platformId=null, journalGroupId=null, journalGroupNameCn=null, journalGroupNameEn=null, type=1, domain=https://castjournals.cast.org.cn/joweb/wswxb/EN, language=EN, createTime=1762149843935, createBy=18614031015, updateTime=1762149925242, updateBy=18614031015, name=微生物学报-英文, tplId=1146101810881728533, title=Acta Microbiologica Sinica, delFlag=0, indexPage=/home, props=[WebsiteProps(id=1192107140455220192, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1192106106018218929, code=articleTextType, value=kx, createTime=1762150090564, updateTime=1762150090564, creator=18614031015, updator=18614031015), WebsiteProps(id=1192107140434248669, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1192106106018218929, code=banner, value=null, createTime=1762150090559, updateTime=1762150090559, creator=18614031015, updator=18614031015), WebsiteProps(id=1192107140476191715, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1192106106018218929, code=grayFlag, value=0, createTime=1762150090569, updateTime=1762150090569, creator=18614031015, updator=18614031015), WebsiteProps(id=1192107140425860060, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1192106106018218929, code=logo, value=https://castjournals.cast.org.cn/joweb/wswxb/EN/file/pic?fileId=FOz4Ks7dC79FYnCEBIlMdw==, createTime=1762150090557, updateTime=1762150090557, creator=18614031015, updator=18614031015), WebsiteProps(id=1192107140484580325, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1192106106018218929, code=minRunFlag, value=0, createTime=1762150090571, updateTime=1762150090571, creator=18614031015, updator=18614031015), WebsiteProps(id=1192107140451025887, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1192106106018218929, code=picServerUrl, value=https://castjournals.cast.org.cn/joweb/wswxb/EN/file/pic, createTime=1762150090563, updateTime=1762150090563, creator=18614031015, updator=18614031015), WebsiteProps(id=1192107140480386020, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1192106106018218929, code=silenceFlag, value=0, createTime=1762150090570, updateTime=1762150090570, creator=18614031015, updator=18614031015), WebsiteProps(id=1192107140442637278, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1192106106018218929, code=staticResourcePath, value=https://castjournals.cast.org.cn/joweb/cast_kjdb_en_623/, createTime=1762150090561, updateTime=1762150090561, creator=18614031015, updator=18614031015), WebsiteProps(id=1192107140463608801, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1192106106018218929, code=themeColor, value=null, createTime=1762150090566, updateTime=1762150090566, creator=18614031015, updator=18614031015), WebsiteProps(id=1192107140467803106, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1192106106018218929, code=themeStyle, value=null, createTime=1762150090567, updateTime=1762150090567, creator=18614031015, updator=18614031015)])], journalTitle=微生物学报, weixinUrl=null, journalUrl=https://actamicro.ijournals.cn, iacademicId=null, status=1, seqNo=null, journalTitleEn=Acta Microbiologica Sinica, journalPhotoCn=tNA7JigLZj/rxynSmzKgDQ==, journalPhotoEn=R/d5eSUu8/o5mAGWCF3M5Q==, journalFirstLetter=A, journalRecommend=null, journalNew=null, journalCollection=null, jcrJf=null, cjcrJf=null, jcrJfStr=null, cjcrJfStr=null, submissionFirstDecision=null, sciSubjectClassification=null, casSubjectClassification=null, citeScore=null, totalCitationFrequency=null, icpCode=null, psCode=null, advertisingLicenseCode=null, copyrightInformation=null, country=null, option=, provinceCode=null, provinceName=null, collectFlag=false), detailUrlCn=https://castjournals.cast.org.cn/joweb/wswxb/CN/10.13343/j.cnki.wsxb.20250750, detailUrlEn=https://castjournals.cast.org.cn/joweb/wswxb/EN/10.13343/j.cnki.wsxb.20250750, pdfUrlCn=https://castjournals.cast.org.cn/joweb/wswxb/CN/PDF/10.13343/j.cnki.wsxb.20250750, pdfUrlEn=https://castjournals.cast.org.cn/joweb/wswxb/EN/PDF/10.13343/j.cnki.wsxb.20250750, aliStartDate=null, aliEndDate=null, collectionFlag=false, citedCount=null, citedUrl=null, reference=null)
收藏切换
群体感应相关调控子对副溶血弧菌磷酸二酯酶GepA的表达调控
收藏切换
PDF下载
刘超 , 李雪 , 罗茜 , 张义全 , 张苗苗 * , 陆仁飞 *
微生物学报 | 研究报告 2026,66(3): 1167-1177
收起
收藏切换
微生物学报 | 研究报告 2026, 66(3): 1167-1177
群体感应相关调控子对副溶血弧菌磷酸二酯酶GepA的表达调控
全屏
刘超, 李雪, 罗茜, 张义全, 张苗苗* , 陆仁飞*
作者信息
  • 南通市第三人民医院，南通大学附属南通第三医院检验科，江苏 南通
Quorum sensing regulators control the expression of phosphodiesterase GepA in Vibrio parahaemolyticus
Chao LIU, Xue LI, Xi LUO, Yiquan ZHANG, Miaomiao ZHANG* , Renfei LU*
Affiliations
  • Department of Clinical Laboratory, Affiliated Nantong Hospital 3 of Nantong University, Nantong Third People’s Hospital, Nantong, Jiangsu, China
出版时间: 2026-03-04 doi: 10.13343/j.cnki.wsxb.20250750
文章导航
收藏切换
摘要
收起

目的 研究群体感应系统(quorum sensing, QS)相关调控子AphA、ToxR和QsvR对副溶血弧菌磷酸二酯酶GepA编码基因gepA表达的调控关系。 方法 提取野生株(wild type, WT)和aphA、toxR、qsvR突变株的总RNA，通过实时定量PCR (quantitative real-time PCR, qPCR)初步分析QS相关调控子对gepA的转录影响；将gepA上游调控区DNA序列克隆入pBBRlux质粒无启动子区的表达生物冷光基因luxCDABE上游，并将lux重组质粒分别导入WT和突变株中，采用lux报告基因融合实验进一步研究QS相关调控子对gepA的调控作用；采用引物延伸(primer extension)法定位gepA的转录起始位点和核心启动子区，并根据引物延伸产物丰度判断QS相关调控子对gepA的调控关系；将gepA的调控区DNA序列克隆入pHRP309质粒中β-半乳糖苷酶基因上游获得LacZ重组质粒，将LacZ重组质粒分别转化入含有pBAD33或pBAD33-qsvR的大肠杆菌EC100 λpir中，采用LacZ报告基因融合实验研究在异体宿主中QS相关调控子对gepA转录的调控方式；PCR扩增gepA上游调控区DNA序列，同时表达并纯化QS相关调控子的His重组蛋白，采用凝胶阻滞实验(electrophoresis mobility shift assay, EMSA)研究QS相关调控子是否直接调控gepA的表达。 结果 低密度条件下，qPCR结果显示ΔaphA和ΔtoxR中gepA的转录水平明显低于WT，表明AphA和ToxR激活gepA的转录；lux报告基因融合实验显示，ΔaphA和ΔtoxR中gepA的启动子区转录活性明显低于WT，进一步表明AphA和ToxR促进gepA的转录；引物延伸结果显示，gepA的转录起始位点位于起始密码子ATG上游第30 bp的A，且其转录活性受到AphA的激活；EMSA结果显示，His-AphA和His-ToxR均不能与gepA的调控区DNA序列结合。高密度条件下，qPCR和引物延伸实验结果均显示QsvR抑制gepA的转录；EMSA实验结果显示，His-QsvR直接结合在gepA的启动子区DNA序列上；双质粒报告基因融合实验显示，QsvR可以抑制EC100 λpir中gepA的启动子区转录活性。 结论 AphA和ToxR间接激活，而QsvR直接抑制gepA的转录。因此，gepA在低密度条件下转录水平较高，高密度时转录水平明显下降。

副溶血弧菌  /  群体感应系统  /  GepA  /  转录调控

Objective To investigate the transcriptional regulation of quorum sensing (QS) regulators AphA, ToxR, and QsvR on the expression of the phosphodiesterase (GepA) gene gepA in Vibrio parahaemolyticus. Methods Total RNAs were extracted from the wild type (WT) and the mutant strains of aphA, toxR, and qsvR. Quantitative real-time PCR (qPCR) was carried out to calculate the transcriptional variation of gepA between WT and mutant strains. The regulatory DNA region of gepA was cloned into the upstream region of promoterless luxCDABEreporter gene in the pBBRlux plasmid. The recombinant plasmid was respectively transferred into the WT and mutant strains. Luminescence assay was used to test the regulatory effect of QS regulators on the expression of gepA. The primer extension assay was employed to detect the transcription start site and the promoter activity of gepA. The effects of QS regulators on gepA were evaluated based on the abundance of primer extension products. The regulatory DNA region of gepA was cloned into the upstream region of lacZ in the pHRP309 plasmid. The LacZ recombinant plasmid was transformed into EC100 λpir harboring pBAD33 or PBAD33-qsvR. Two-plasmid LacZ reporter assay was conducted to investigate the regulatory effects of QS regulators on the transcription of gepA in EC100 λpir. The regulatory DNA region of gepA was amplified by PCR, and the His recombinant proteins of QS regulators were purified. The electrophoretic mobility shift assay (EMSA) was performed to investigate whether QS regulators directly regulated the expression of gepA. Results At low cell density, the qPCR results showed that expression of gepA in ΔaphA and ΔtoxR were significantly lower than that in WT, indicating that AphA and ToxR activated the transcription of gepA. The luminescence assay showed that the transcriptional activity of the promoter region of gepA in ΔaphA and ΔtoxR was significantly lower than that in WT, further indicating that AphA and ToxR promoted the transcription of gepA. The primer extension assay detected that the transcription start site of gepA was located at the A nucleotide 30 bp upstream of the start codon ATG, and its transcriptional activity was activated by AphA. The EMSA result indicated that His-AphA and His-ToxR were unable to bind the promoter DNA region of gepA. At high cell density, both the qPCR and primer extension assay indicated that QsvR inhibited the transcription of gepA. The EMSA result demonstrated that His-QsvR directly bound to the promoter DNA region of gepA. Two-plasmid lacZ reporter assay demonstrated that QsvR inhibited the transcriptional activity of the promoter region of gepA in EC100 λpir. Conclusion AphA and ToxR indirectly activate while QsvR directly inhibits the transcription of gepA. Therefore, the transcription level of gepA is higher at low cell density and significantly decreases at high cell density.

Vibrio parahaemolyticus  /  quorum sensing  /  GepA  /  transcriptional regulation
刘超, 李雪, 罗茜, 张义全, 张苗苗, 陆仁飞. 群体感应相关调控子对副溶血弧菌磷酸二酯酶GepA的表达调控. 微生物学报, 2026 , 66 (3) : 1167 -1177 . DOI: 10.13343/j.cnki.wsxb.20250750
Chao LIU, Xue LI, Xi LUO, Yiquan ZHANG, Miaomiao ZHANG, Renfei LU. Quorum sensing regulators control the expression of phosphodiesterase GepA in Vibrio parahaemolyticus[J]. Acta Microbiologica Sinica, 2026 , 66 (3) : 1167 -1177 . DOI: 10.13343/j.cnki.wsxb.20250750
正文
收起
副溶血弧菌(Vibrio parahaemolyticus)是弧菌属的一种革兰氏阴性嗜盐菌，可引发海产品和人类感染，是世界范围内海产品相关性食物中毒的重要致病菌[1]。副溶血弧菌可表达多种毒力因子，如耐热直接溶血素(thermostable direct hemolysin, TDH)、TDH相关溶血素(TDH-related hemolysin, TRH)、III型分泌系统(type III secretion system, T3SS)、VI型分泌系统(type VI secretion system, T6SS)等，这些因子参与细菌致病过程[1-3]。副溶血弧菌具有较强的表面生物膜形成能力，能够逃避宿主免疫反应，并抵抗外界不利生存环境[4]。成熟生物膜的形成依赖于胞外多糖、Ⅳ型菌毛、荚膜多糖、鞭毛等特殊物质和结构[4-5]。此外，生物膜形成过程受到一些信号通路，如群体感应系统(quorum sensing, QS)和环二鸟苷酸(c-di-GMP)等的严格调控[4]。
c-di-GMP是一种在细菌中广泛存在的第二信使，由含有GGDEF结构域的鸟苷酸环化酶(diguanylate cyclase, DGC)催化2分子GTP合成，可被含有EAL或HD-GYP结构域的磷酸二酯酶(phosphodiesterase, PDE)降解为2分子GMP或线性pGpG[6-7]。c-di-GMP可调控细菌的毒力因子表达、运动能力、生物膜形成和细胞周期等多种生物学行为[8]。细菌体内c-di-GMP浓度升高有助于生物膜形成，但会抑制运动能力和毒力因子表达[8]。副溶血弧菌RIMD2210633可编码63种含有GGDEF和/或EAL结构域的蛋白质，目前仅有少数蛋白质的功能得到证实，其中ScrC、ScrG、TpdA和VopY等可促进细菌运动，但降低生物膜形成能力；而GefA、ScrO、ScrJ、ScrL、GefB和VPA0198等则抑制细菌运动，但提高生物膜形成能力[9-17]。
群体感应系统是指细菌响应周围菌体密度变化，合成并分泌自诱导因子，通过一系列信号传递，最终调节菌体基因表达和生物学行为的过程[18]。在副溶血弧菌中，AphA和OpaR分别在低密度和高密度条件下发挥主要调控作用，是群体感应系统的核心调控子[18]。低密度条件下AphA发挥调控作用，促进胞外多糖(exopolysaccharide, EPS)的产生和T3SS1的表达，抑制TDH和T3SS2的表达，从而增强副溶血弧菌的生物膜形成和侵袭能力[19-20]；QsvR是AraC家族转录调控子，由操纵子vpa0607-qsvR编码，VPA0607蛋白具有核糖核酸酶Ⅱ活性，能在转录后抑制QsvR的表达[21]。AphA激活vpa0607-qsvR的转录，而OpaR抑制其转录，因此QsvR在高密度时表达水平最高，从而与OpaR共同发挥调控作用，抑制EPS的产生和T3SS1的表达，激活TDH和T3SS2的表达[20-22]。此外，QsvR对极鞭毛和T6SS1相关基因也具有抑制作用[20,23]。ToxR是一种膜结合调控蛋白，其自身基因的转录受到AphA的抑制，在低密度条件下转录水平最高[24]。ToxR与AphA和OpaR协同作用抑制T6SS1的表达，同时抑制T3SS1的表达，促进T3SS2和TDH2的表达以及生物膜的形成[24-26]。可见，QsvR和ToxR与AphA和OpaR组成了复杂的调控网络，使QsvR和ToxR参与到群体感应系统的调控机制中。
群体感应系统相关调控子均参与调控c-di-GMP的代谢，AphA促进c-di-GMP的合成，而ToxR、QsvR和OpaR抑制其合成，但其具体调控机制尚未完全阐明[22,27-28]。近期研究发现GepA (VP0117)同时含有GGDEF和EAL结构域，但仅具有PDE活性，具有降解c-di-GMP、抑制生物膜形成但促进运动能力的作用[29]。同时也有研究表明，OpaR直接抑制gepA的转录，其他群体感应系统相关调控子对gepA的转录是否有调控作用尚不清楚[30]。因此，本研究旨在探究AphA、ToxR和QsvR对gepA的转录调控机制。
1 材料与方法
收起
1.1 材料
1.1.1 菌株
副溶血弧菌RIMD2210633 (野生型，WT)及其aphA非极性突变株(ΔaphA)、toxR非极性突变株(ΔtoxR)、qsvR非极性突变株(ΔqsvR)、重组蛋白表达菌等由南通大学附属南通第三医院检验科保存[20,24]。
1.1.2 主要试剂和仪器
HI培养基(2.5% Bacto heart infusion)，BD Bioscience公司；TRIzol Reagent，Invitrogen公司；Primer Extension System、β-galactosidase Enzyme Assay System，Promega公司；AccuPower and Top DNA Sequencing Kit，Bioneer公司；2×Taq PCR MasterMix、SuperReal荧光定量预混试剂彩色版(SYBR Green)、FastKing一步法除基因组cDNA第一链合成预混试剂、普通DNA产物纯化试剂盒，天根生化科技(北京)有限公司。
恒温摇床、恒温细菌培养箱、紫外分光光度计和酶标仪，ThermoFisher Scientific公司；实时荧光定量PCR仪，Bio-Rad公司；凝胶成像系统，上海勤翔科学仪器有限公司。
1.2 细菌培养
取10 μL甘油菌种接种于5 mL的HI肉汤中，37 ℃、200 r/min培养12 h。将培养产物1:50接种至5 mL新鲜HI肉汤中，培养至OD600约为1.4。再将培养产物1:100接种至5 mL新鲜HI肉汤中，培养至OD600约为0.2 (低密度)或OD600约为0.8 (高密度)，收集菌体[20]。在某些情况下还需添加阿拉伯糖和抗生素，其工作浓度如下：阿拉伯糖为0.1%、氯霉素为20 μg/mL、庆大霉素为100 μg/mL。
1.3 实时定量PCR
采用TRIzol试剂提取细菌的总RNA。分别取1 μg的总RNA，利用FastKing一步法除基因组cDNA第一链合成预混试剂盒制备cDNA，进而采用SuperReal荧光定量预混试剂彩色版(SYBR Green)进行实时定量PCR (quantitative real-time PCR, qPCR)分析。以16S rRNA基因的表达量为内参，采用经典的2 -ΔΔCt法对gepA的转录水平进行相对定量[31]。所用引物列于表1中。
1.4 lux 报告基因融合实验(luminescence assay)
将gepA的上游调控区DNA序列克隆入pBBRlux质粒中无启动子区的自发光基因LuxCDABE上游，构建Lux-gepA重组质粒，并将其分别导入WT和突变株中，获得lux实验菌株。按照1.2节方法培养lux实验菌株，测定不同菌株的冷光值(Lux)，并计算相对平均冷光单位(relative light unit, RLU)，如公式(1)所示。
RLU=Lux/OD600[23]
1.5 引物延伸(primer extension)
将能与gepA mRNA互补的特异性引物(表1)的5′-末端用[γ-32P]-ATP (5 000 Ci/mmol)进行放射性标记。分别以等量的WT和突变株总RNA为模板，利用Primer Extension System进行引物延伸实验，将gepA的mRNA逆转录成cDNA。将逆转录产物与Sanger测序条带进行6%聚丙烯酰胺变性胶凝胶电泳，-20 ℃放射自显影后分析结果[20]。
1.6 双质粒报告基因融合实验
将gepA上游调控区DNA序列克隆入pHRP309质粒中无启动子的β-半乳糖苷酶基因上游，构建LacZ质粒，并将其分别转入携带pBAD33和pBAD33-qsvR重组质粒的大肠杆菌100 λpir (EC100 λpir)中，获得双质粒报告基因融合实验菌株。采用LB肉汤培养基(1.0%胰蛋白胨、0.5%酵母提取物和1.0% NaCl)在37 ℃下培养双质粒报告基因融合实验菌株，收集对数中期(OD600约为1.2)的细菌培养产物。采用β-galactosidase Enzyme Assay System检测不同菌株中的β-半乳糖苷酶活性。β-半乳糖苷酶活性用Miller units表示，其计算如公式(2)所示。
β-半乳糖苷酶活性=106×[(OD420-1.75×OD550)]/(T×V×OD600)
式中：T表示反应时间(min)，V表示反应体系的体积(μL)。通过比较Miller units数值差异，可以判断gepA启动子区在不同菌株中的活性[32]。
1.7 凝胶阻滞实验(electrophoresis mobility shift assay, EMSA)
PCR扩增gepA上游调控区DNA序列(引物见表1)，并用T4多聚核苷酸激酶和[γ-32P]-ATP (5 000 Ci/mmol)对其5′-末端进行放射性标记，制备EMSA探针。表达并纯化His重组蛋白，并将不同浓度的His重组蛋白与EMSA探针在10 μL结合体系(1 mmol/L MgCl2，0.5 mmol/L EDTA，0.5 mmol/L DTT，50 mmol/L NaCl，10 mmol/L pH 7.5的Tris-HCl，0.05 mg/mL鲑鱼精DNA，EMSA探针)中共孵育20 min (室温条件下)，而后进行6%非变性聚丙烯酰胺凝胶电泳，-20 ℃放射自显影后分析结果[26]。
1.8 统计学分析方法
引物延伸和EMSA至少重复2次，并获得相似或相同的实验结果。qPCR和LacZ报告基因融合实验至少重复3次，每次至少包含3个生物学重复，结果用平均值±标准差(standard deviation, SD)表示，利用双尾t检验进行统计学分析，以P<0.01为依据，认为具有显著性差异。
2 结果与分析
收起
2.1 AphA正调控 gepA 的转录
提取低密度条件下细菌的总RNA，采用qPCR实验研究AphA对gepA的调控关系。结果显示(图1A)，在ΔaphA中检测到的gepA mRNA丰度明显低于在WT中检测到的，表明AphA对gepA的转录具有促进作用。构建Lux-gepA重组质粒，并将其分别导入WT和ΔaphA中，采用lux报告基因融合实验进一步研究AphA对gepA的调控关系。与WT相比，在ΔaphA中检测到的RLU明显降低，表明AphA正调控gepA的转录(图1B)。引物延伸实验显示(图1C)，gepA的转录起始位点位于起始密码子ATG上游第30 bp的A处，且在ΔaphA中检测到的引物延伸产物丰度显著低于在WT中检测到的，这进一步表明AphA正调控gepA的转录。PCR扩增gepA的调控区DNA序列，并用[γ-32P]-ATP (5 000 Ci/mmol)对其5′-末端进行放射性标记，采用EMSA实验验证His-AphA与gepA的调控区DNA序列是否具有结合作用。如图1D所示，无论加入依次递增的His-AphA (第1-4泳道)，还是额外加入未标记的核酸探针作为竞争DNA (第5泳道)，或者加入无关蛋白(第7泳道)时均未出现阻滞条带，表明His-AphA不能与gepA的调控区DNA结合，因此AphA是间接调控gepA转录的。16S rRNA作为阴性对照，不能与His-AphA结合[20]。上述结果表明，AphA间接正调控gepA的转录。
2.2 ToxR正调控 gepA 的转录
在低密度条件下qPCR结果显示(图2A)，在ΔtoxR中检测到的gepA mRNA丰度明显低于WT，这表明ToxR促进gepA的转录。采用lux报告基因融合实验研究ToxR对gepA的调控关系(图2B)，与WT相比，在ΔtoxR中检测到的RLU明显降低，表明ToxR正调控gepA的转录。EMSA结果显示(图2C)，His-ToxR与gepA上游调控区序列未出现阻滞条带，表明ToxR间接调控gepA的转录。16S rDNA作为阴性对照，不能与His-ToxR结合[24]。上述结果表明，ToxR间接正调控gepA的转录。
2.3 QsvR负调控 gepA 的转录
高密度条件下qPCR结果显示(图3A)，在ΔqsvR中检测到的gepA mRNA丰度明显高于在WT中检测到的，说明QsvR抑制gepA的转录。引物延伸实验显示(图3B)，与上述结果一致，gepA转录起始位点仍位于起始密码子ATG上游第30 bp的A处，且在ΔqsvR中检测到的引物延伸产物丰度显著高于在WT中检测到的，这进一步表明QsvR负调控gepA的转录。PCR扩增gepA的调控区DNA序列，采用EMSA实验验证在体外条件下His-QsvR与gepA的调控区DNA是否具有结合作用。如图3C所示，随着His-QsvR用量依次递增(第2-7泳道)，与gepA的调控区DNA序列的阻滞条带强度明显增强，表明His-QsvR能与gepA的调控区DNA结合，且呈浓度依赖性。vp1687作为阴性对照，其上游调控区DNA序列不能与His-QsvR结合，未出现阻滞条带。由于在体外条件下，QsvR可以和gepA的调控区DNA结合，因此将gepA的LacZ重组质粒分别转入携带pBAD33和pBAD33重组质粒的大肠杆菌100 λpir中，采用LacZ实验研究异源宿主中表达的QsvR是否能调控靶基因启动子区活性。如图3D所示，在EC100/pBAD33-qsvR中检测到的β-半乳糖苷酶活性显著低于在EC100/pBAD33中检测到的，这说明在异源宿主中QsvR的过表达也可以抑制gepA的启动子区活性。由于大肠杆菌和副溶血弧菌的遗传背景差异较大，图3D的结果也说明QsvR对gepA的启动子区DNA序列具有直接的结合活性。上述结果表明，QsvR直接负调控gepA的转录。
2.4 gepA 呈密度依赖性转录
采用lux报告基因融合实验测定gepA在不同生长时期的转录情况(图4)。结果表明，在OD600值为0.02-0.30的范围内，随着细胞密度的增加，gepA启动子区活性显著升高，在OD600值为0.3-0.4时达到最高水平，OD600值大于0.4时下降。这表明gepA的转录水平与菌体密度相关。
3 讨论与结论
收起
细菌内c-di-GMP的合成与降解是一个受到严格调控的过程。在副溶血弧菌中QS核心调控子及其相关调控子均可参与调控c-di-GMP代谢，AphA间接抑制scrABC和scrG的转录，从而促进c-di-GMP的合成和生物膜形成[27]；OpaR和QsvR协同作用抑制scrABC而激活scrG的转录，进而抑制c-di-GMP的合成和生物膜形成[22]；此外，OpaR也直接抑制VPA0198、VPA1176、VP0699和gepA，并间接激活VP2979的转录[30]。ToxR通过直接抑制scrA和scrG的转录，以及间接抑制vpa0198的转录来抑制c-di-GMP的合成[28]。H-NS对scrA、gepA、VPA0198、VPA1176、VP0699、scrG和VP2979的转录也具有直接抑制作用，从而降低胞内c-di-GMP浓度，抑制生物膜形成[33]。此外，低盐生长条件抑制scrABC和scrG的转录，进而抑制c-di-GMP的降解，促进生物膜形成[34]。
c-di-GMP的合成和降解分别受鸟苷酸环化酶和磷酸二酯酶的催化，这2种酶分别具有GGEEF结构域和EAL结构域[35]。菌体内c-di-GMP浓度升高有利于细菌生物膜的形成，但会抑制细菌的运动能力[35]。近期研究发现，具有GGDEF和EAL结构域的GepA在菌体内具有降解c-di-GMP、抑制生物膜形成以及促进运动能力的功能[29]。本研究主要探究gepA的转录调控机制，结果表明在低密度条件下AphA和ToxR间接激活gepA的转录；而在高密度条件下QsvR直接抑制gepA的转录(图5)。先前研究发现，OpaR也能够直接抑制gepA的转录[30]。gepA的转录水平与菌体密度密切相关，低密度时gepA启动子区活性随着细胞密度的增加显著升高，当OD600值为0.3-0.4时达到最高水平，而当OD600值大于0.4时则逐渐下降(图4)。
前期研究表明，QsvR直接抑制toxR和aphA的转录，同时直接激活opaR的转录；此外，AphA间接抑制qsvR的转录，而OpaR直接激活其转录[30,32]。AphA间接抑制toxR的转录[24]。由此可见，副溶血弧菌gepA的表达受QS系统(AphA和OpaR)及其他相关调控子(QsvR和ToxR)组成的调控网络紧密调控。然而，gepA在转录水平最高的低细菌密度条件下如何发挥作用，以及gepA与QsvR的结合位点还有待进一步实验研究。
基金
收起
  • 南通市自然科学基金(JCZ2025004)
文献
收起
参考文献 引证文献
排序方式：
[1]
ZAKARIA NH, RAHIM NDE ABD, ROSILAN NF, SUNG YY, WAIHO K, HARUN S, ZAINAL ABIDIN RA, AFIQAH-ALENG N. Global landscape of Vibrio parahaemolyticus research: a bibliometric analysis[J]. World Journal of Microbiology and Biotechnology, 2025, 41(2): 45.
[2]
SHIMOHATA T, TAKAHASHI A. Diarrhea induced by infection of Vibrio parahaemolyticus [J]. Journal of Medical Investigation, 2010, 57(3/4): 179-182.
[3]
HIYOSHI H, KODAMA T, IIDA T, HONDA T. Contribution of Vibrio parahaemolyticus virulence factors to cytotoxicity, enterotoxicity, and lethality in mice[J]. Infection and Immunity, 2010, 78(4): 1772-1780.
[4]
ASHIKUR RAHMAN M, AKTER S, ASHRAFUDOULLA M, ANAMUL HASAN CHOWDHURY M, UDDIN MAHAMUD AGMS, PARK SH, HA SD. Insights into the mechanisms and key factors influencing biofilm formation by Aeromonas hydrophila in the food industry: a comprehensive review and bibliometric analysis[J]. Food Research International, 2024, 175: 113671.
[5]
ASHRAFUDOULLA M, MIZAN MFR, PARK SH, HA SD. Current and future perspectives for controlling Vibrio biofilms in the seafood industry: a comprehensive review[J]. Critical Reviews in Food Science and Nutrition, 2021, 61(11): 1827-1851.
[6]
JENAL U, REINDERS A, LORI C. Cyclic di-GMP: second messenger extraordinaire[J]. Nature Reviews Microbiology, 2017, 15(5): 271-284.
[7]
KALIA D, MEREY G, NAKAYAMA S, ZHENG Y, ZHOU J, LUO YL, GUO M, ROEMBKE BT, SINTIM HO. Nucleotide, c-di-GMP, c-di-AMP, cGMP, cAMP, (p)ppGpp signaling in bacteria and implications in pathogenesis[J]. Chemical Society Reviews, 2013, 42(1): 305-341.
[8]
HOMMA M, KOJIMA S. Roles of the second messenger c-di-GMP in bacteria: focusing on the topics of flagellar regulation and Vibrio spp.[J]. Genes to Cells, 2022, 27(3): 157-172.
[9]
TRIMBLE MJ, McCARTER LL. Bis-(3′-5′)-cyclic dimeric GMP-linked quorum sensing controls swarming in Vibrio parahaemolyticus [J]. Proceedings of the National Academy of Sciences of the United States of America, 2011, 108(44): 18079-18084.
[10]
KIM YK, McCARTER LL. ScrG, a GGDEF-EAL protein, participates in regulating swarming and sticking in Vibrio parahaemolyticus [J]. Journal of Bacteriology, 2007, 189(11): 4094-4107.
[11]
MARTÍNEZ-MÉNDEZ R, CAMACHO-HERNÁNDEZ DA, SULVARÁN-GUEL E, ZAMORANO-SÁNCHEZ D. A trigger phosphodiesterase modulates the global c-di-GMP pool, motility, and biofilm formation in Vibrio parahaemolyticus [J]. Journal of Bacteriology, 2021, 203(13): e0004621.
[12]
WU X, ZHOU LT, YE C, ZHA ZZ, LI CC, FENG C, ZHANG Y, JIN Q, PAN JY. Destruction of self-derived PAMP via T3SS2 effector VopY to subvert PAMP-triggered immunity mediates Vibrio parahaemolyticus pathogenicity[J]. Cell Reports, 2023, 42(11): 113456.
[13]
ZHONG XJ, LU Z, WANG F, YAO N, SHI MT, YANG MH. Characterization of GefA, a GGEEF domain-containing protein that modulates Vibrio parahaemolyticus motility, biofilm formation, and virulence[J]. Applied and Environmental Microbiology, 2022, 88(6): e0223921.
[14]
KIMBROUGH JH, CRIBBS JT, McCARTER LL. Homologous c-di-GMP-binding scr transcription factors orchestrate biofilm development in Vibrio parahaemolyticus [J]. Journal of Bacteriology, 2020, 202(6): e00723-19.
[15]
KIMBROUGH JH, McCARTER LL. Identification of three new GGDEF and EAL domain-containing proteins participating in the scr surface colonization regulatory network in Vibrio parahaemolyticus [J]. Journal of Bacteriology, 2021, 203(4): e00409-20.
[16]
ZHOU YN, CHANG JY, ZHANG MM, LI X, LUO X, LI WP, TIAN ZK, ZHANG N, NI B, ZHANG YQ, LU RF. GefB, a GGDEF domain-containing protein, affects motility and biofilm formation of Vibrio parahaemolyticus and is regulated by quorum sensing regulators[J]. Gene, 2025, 933: 148968.
[17]
ZHOU YN, CHANG JY, ZHANG MM, LI X, YANG WH, HU LF, ZHOU DS, NI B, LU RF, ZHANG YQ. VPA0198, a GGDEF domain-containing protein, affects the motility and biofilm formation of Vibrio parahaemolyticus and is regulated by quorum sensing associated regulators[J]. Microbial Pathogenesis, 2024, 195: 106882.
[18]
GODE-POTRATZ CJ, McCARTER LL. Quorum sensing and silencing in Vibrio parahaemolyticus [J]. Journal of Bacteriology, 2011, 193(16): 4224-4237.
[19]
CHEN L, ZHANG MM, LI X, WU QM, XUE XF, ZHANG TT, LU RF, ZHANG YQ. AphA directly activates the transcription of polysaccharide biosynthesis gene scvE in Vibrio parahaemolyticus [J]. Gene, 2023, 851: 146980.
[20]
ZHANG YQ, HU LH, QIU Y, OSEI-ADJEI G, TANG H, ZHANG Y, ZHANG R, SHENG XM, XU SG, YANG WH, YANG HY, YIN Z, YANG RF, HUANG XX, ZHOU DS. QsvR integrates into quorum sensing circuit to control Vibrio parahaemolyticus virulence[J]. Environmental Microbiology, 2019, 21(3): 1054-1067.
[21]
ZHANG YQ, XUE XF, SUN FJ, LI X, ZHANG MM, WU QM, ZHANG TT, LUO X, LU RF. Quorum sensing and QsvR tightly control the transcription of vpa0607 encoding an active RNase II-type protein in Vibrio parahaemolyticus [J]. Frontiers in Microbiology, 2023, 14: 1123524.
[22]
ZHANG MM, XUE XF, LI X, WU QM, ZHANG TT, YANG WH, HU LF, ZHOU DS, LU RF, ZHANG YQ. QsvR and OpaR coordinately repress biofilm formation by Vibrio parahaemolyticus [J]. Frontiers in Microbiology, 2023, 14: 1079653.
[23]
ZHANG MM, XUE XF, LI X, LUO X, WU QM, ZHANG TT, YANG WH, HU LF, ZHOU DS, LU RF, ZHANG YQ. QsvR represses the transcription of polar flagellum genes in Vibrio parahaemolyticus [J]. Microbial Pathogenesis, 2023, 174: 105947.
[24]
ZHANG YQ, HU LF, OSEI-ADJEI G, ZHANG Y, YANG WH, YIN Z, LU RY, SHENG XM, YANG RF, HUANG XX, ZHOU DS. Autoregulation of ToxR and its regulatory actions on major virulence gene loci in Vibrio parahaemolyticus [J]. Frontiers in Cellular and Infection Microbiology, 2018, 8: 291.
[25]
ZHANG YQ, GAO H, OSEI-ADJEI G, ZHANG Y, YANG WH, YANG HY, YIN Z, HUANG XX, ZHOU DS. Transcriptional regulation of the type VI secretion system 1 genes by quorum sensing and ToxR in Vibrio parahaemolyticus [J]. Frontiers in Microbiology, 2017, 8: 2005.
[26]
OSEI-ADJEI G, GAO H, ZHANG Y, ZHANG LY, YANG WH, YANG HY, YIN Z, HUANG XX, ZHANG YQ, ZHOU DS. Regulatory actions of ToxR and CalR on their own genes and type III secretion system 1 in Vibrio parahaemolyticus [J]. Oncotarget, 2017, 8(39): 65809-65822.
[27]
黄倩, 张义全, 胡小许, 王丽, 杨瑞馥, 钟青萍, 周冬生, 黎晓敏. AphA蛋白促进副溶血弧菌c-di-GMP合成和生物膜形成[J]. 微生物学报, 2014, 54(5): 525-531.
HUANG Q, ZHANG YQ, HU XX, WANG L, YANG RF, ZHONG QP, ZHOU DS, LI XM. AphA is an activator of c-di-GMP synthesis and biofilm formation in Vibrio parahaemolyticus [J]. Acta Microbiologica Sinica, 2014, 54(5): 525-531 (in Chinese).
[28]
张苗苗, 薛星帆, 孙君芳, 吴齐敏, 李雪, 周冬生, 倪斌, 陆仁飞, 张义全. ToxR负调控副溶血弧菌中c-di-GMP的合成[J]. 生物工程学报, 2022, 38(12): 4719-4730.
ZHANG MM, XUE XF, SUN JF, WU QM, LI X, ZHOU DS, NI B, LU RF, ZHANG YQ. ToxR represses the synthesis of c-di-GMP in Vibrio parahaemolyticus [J]. Chinese Journal of Biotechnology, 2022, 38(12): 4719-4730 (in Chinese).
[29]
ZHANG MM, ZHU YR, LI X, LUO X, SUN H, XIONG SH, LU RF, ZHANG YQ. GepA, a GGDEF-EAL protein, regulates biofilm formation and swimming motility in Vibrio parahaemolyticus [J]. Archives of Microbiology, 2025, 207(5): 99.
[30]
ZHANG YQ, QIU Y, GAO H, SUN JF, LI X, ZHANG MM, XUE XF, YANG WH, NI B, HU LF, YIN Z, LU RF, ZHOU DS. OpaR controls the metabolism of c-di-GMP in Vibrio parahaemolyticus [J]. Frontiers in Microbiology, 2021, 12: 676436.
[31]
MA YJ, SUN XH, XU XY, ZHAO Y, PAN YJ, HWANG CA, WU VCH. Investigation of reference genes in Vibrio parahaemolyticus for gene expression analysis using quantitative RT-PCR[J]. PLoS One, 2015, 10(12): e0144362.
[32]
ZHANG YQ, QIU Y, XUE XF, ZHANG MM, SUN JF, LI X, HU LF, YIN Z, YANG WH, LU RF, ZHOU DS. Transcriptional regulation of the virulence genes and the biofilm formation associated operons in Vibrio parahaemolyticus [J]. Gut Pathogens, 2021, 13(1): 15.
[33]
XUE XF, ZHNAG MM, SUN JF, LI X, WU QM, YIN Z, YANG WH, NI B, HU LF, ZHOU DS, LU RF, ZHANG YQ. H-NS represses biofilm formation and c-di-GMP synthesis in Vibrio parahaemolyticus [J]. Biomedical and Environmental Sciences, 2022, 35(9): 821-829.
[34]
LI X, SUN JF, ZHANG MM, XUE XF, WU QM, YANG WH, YIN Z, ZHOU DS, LU RF, ZHANG YQ. The effect of salinity on biofilm formation and c-di-GMP production in Vibrio parahaemolyticus [J]. Current Microbiology, 2021, 79(1): 25.
[35]
LIU C, SHI R, JENSEN MS, ZHU JR, LIU JW, LIU XB, SUN D, LIU WJ. The global regulation of c-di-GMP and cAMP in bacteria[J]. mLife, 2024, 3(1): 42-56.
2026年第66卷第3期
PDF下载
133
55
引用本文
BibTeX
文章信息
doi: 10.13343/j.cnki.wsxb.20250750
  • 接收时间：2025-10-07
  • 首发时间：2026-03-12
  • 出版时间：2026-03-04
补充材料
相关文章
文章信息
作者
出版历史
  • 收稿日期：2025-10-07
  • 录用日期：2025-12-09
基金
Natural Science Foundation of Nantong(JCZ2025004)
南通市自然科学基金(JCZ2025004)
作者信息
    南通市第三人民医院，南通大学附属南通第三医院检验科，江苏 南通

通讯作者:

通讯作者:

参考文献
分享链接
https://castjournals.cast.org.cn/joweb/wswxb/CN/10.13343/j.cnki.wsxb.20250750
分享至
全文二维码

扫描看全文

引用本文
BibTeX
本文的引用情况
2种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
关闭全屏
相关链接
论文
最新文章
热读文章
热点专题
内参
蓝皮书
产业发展报告
传播力报告
期刊分级目录
资讯
学术新闻
行业新闻
学术会议
行业会议
关于
关于我们
联系我们
北京市海淀区学院南路86号
100081
010-62199257
qkjq@cast.org.cn

中国科学技术协会版权所有 京ICP 备10216604号-4 海淀分局备案 1101084647
科技导报社主办 中国科协信息中心技术支持