Article(id=1304414961585901858, tenantId=1146029695717560320, journalId=1302319053441957962, issueId=1304414955046985824, articleNumber=null, orderNo=null, doi=10.7501/j.issn.0253-2670.2026.07.023, pmid=null, cstr=null, oa=null, hot=0, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1759334400000, receivedDateStr=2025-10-02, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1788926361700, onlineDateStr=2026-09-09, pubDate=null, pubDateStr=null, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1788926361700, onlineIssueDateStr=2026-09-09, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1788926361700, creator=13701087609, updateTime=1788926361700, updator=13701087609, issue=Issue{id=1304414955046985824, tenantId=1146029695717560320, journalId=1302319053441957962, year='2026', volume='57', issue='7', pageStart='2445', pageEnd='2876', issueExtLink='null', onlineDate='null', pubDate='1775923200000', pubDateStr='2026-04-12', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1788926360140, creator='13701087609', updateTime=1788926711174, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1304416427457409395, tenantId=1146029695717560320, journalId=1302319053441957962, issueId=1304414955046985824, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1304416427457409396, tenantId=1146029695717560320, journalId=1302319053441957962, issueId=1304414955046985824, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=2718, endPage=2729, ext={EN=ArticleExt(id=1304414961892086052, articleId=1304414961585901858, tenantId=1146029695717560320, journalId=1302319053441957962, language=EN, title=Gene cloning and expression analysis of cytochrome P450 family gene CaCYP2 from Centella asiatica, columnId=null, journalTitle=Chinese Traditional and Herbal Drugs, columnName=null, runingTitle=null, highlight=null, articleAbstract=Objective To clone the cytochrome P450 gene (CaCYP2) involved in C-6β hydroxylation modification of triterpenoid saponins in Centella asiatica, systematically analyze its molecular characteristics, tissue expression pattern and subcellular localization, and achieve prokaryotic soluble expression, so as to provide experimental basis for clarifying its function in triterpenoid biosynthesis. Methods The full-length sequence of CaCYP2 was cloned from C. asiatica cDNA and subjected to bioinformatics analysis. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was performed to detect the gene specific expression in different tissues. Seamless cloning technology was employed to construct yeast two-hybrid vector, prokaryotic expression vector and subcellular localization vector for self-activation detection, recombinant protein expression and tobacco transient localization analysis, respectively. Results The CaCYP2 gene was successfully cloned. Its open reading frame (ORF) was 1 443 bp, encoding 480 amino acids, and it belonged to the CYP716 family, hence named CYP716E116. The predicted protein had a relative molecular mass of 54 430 and a theoretical isoelectric point of 8.98, showing hydrophilicity. It contained a typical CYP90-like conserved domain and one transmembrane domain. Subcellular localization results indicated that this protein was localized in the endoplasmic reticulum. qRT-PCR analysis showed that the expression level of CaCYP2 was highest in the leaves, which was 6.2 times higher than that in the roots (P < 0.000 1), showing significant differences compared to other tissues. Prokaryotic expression results indicated that a soluble recombinant protein with a relative molecular mass of approximately 91 300 (including a 40 300 tag protein) was obtained after isopropyl β-D-thiogalactoside (IPTG) induction. The yeast two-hybrid experiment confirmed that the yeast two-hybrid vector had no self-activating activity and could be used for subsequent screening of interacting proteins. Conclusion This study successfully cloned the CaCYP2 gene and characterized its molecular structure, tissue-specific expression pattern, and endoplasmic reticulum localization. It also confirmed that the protein can be expressed solubly in a prokaryotic system. These findings lay an important foundation for further investigation into its catalytic function and regulatory role in the biosynthetic pathway of triterpenoid saponins in C. asiatica., authors=LIU Huanying, LEI Ming, XIAO Xiao, LI Linxuan, LI Cui, HUANG Yuan, HUO Juan, WAN Lingyun, PAN Limei, ZHANG Zhanjiang, authorsList=LIU Huanying, LEI Ming, XIAO Xiao, LI Linxuan, LI Cui, HUANG Yuan, HUO Juan, WAN Lingyun, PAN Limei, ZHANG Zhanjiang, authorCompany=null, correspAuthors=null, authorNote=null, correspAuthorsNote=null, 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, fund=null), CN=ArticleExt(id=1304414961804005667, articleId=1304414961585901858, tenantId=1146029695717560320, journalId=1302319053441957962, language=CN, title=积雪草细胞色素P450家族CaCYP2基因克隆及表达分析, columnId=1304140203967410414, journalTitle=中草药, columnName=药材与资源, runingTitle=null, highlight=null, articleAbstract=目的 克隆积雪草Centella asiatica中参与三萜皂苷C-6β羟基化修饰的细胞色素P450基因（CaCYP2），系统解析其分子特征、组织表达模式及亚细胞定位，并实现原核可溶性表达，为阐明其在三萜生物合成中的功能提供实验基础。方法 从积雪草cDNA中克隆CaCYP2全长序列，并进行生物信息学分析，采用实时荧光定量PCR检测其组织特异性表达情况；利用无缝克隆技术构建酵母双杂交载体、原核表达载体及亚细胞定位载体，分别进行自激活检测、重组蛋白表达及烟草瞬时定位分析。结果 成功克隆CaCYP2基因，其开放阅读框（open reading frame，ORF）1 443 bp，编码480个氨基酸，属于CYP716家族，命名为CYP716E116。预测蛋白相对分子质量为54 425.09，理论等电点8.98，呈亲水性，含典型CYP90-like保守结构域及1个跨膜结构域。亚细胞定位结果显示，该蛋白定位于内质网。qRT-PCR分析表明，CaCYP2在叶片中表达量最高，是根中表达量的6.2倍（P＜0.000 1），与其他组织存在显著差异；原核表达结果表明，经异丙基硫代半乳糖苷（isopropylβ-D-thiogalactoside，IPTG）诱导后获得相对分子质量约为91 300的可溶性重组蛋白（含40 300的标签蛋白）；酵母双杂交实验证实，酵母双杂交载体无自激活活性，可用于后续互作蛋白筛选。结论 成功克隆CaCYP2基因，明确其分子结构特征、组织表达特异性及内质网定位，证实其可在原核系统中进行可溶性表达，为深入研究其催化功能及在积雪草三萜皂苷生物合成途径中的调控作用奠定重要的基础。, authors=刘焕英^1,2, 雷明^2,3, 肖潇², 李林轩², 李翠^2,3, 黄媛², 霍娟², 万凌云², 潘丽梅², 张占江^1,2,3, authorsList=刘焕英, 雷明, 肖潇, 李林轩, 李翠, 黄媛, 霍娟, 万凌云, 潘丽梅, 张占江, authorCompany=1 广西中医药大学药学院, 广西 南宁 530200;
2 广西壮族自治区药用植物园/广西道地药材高品质形成与应用重点实验室/国家中医药传承创新中心, 广西 南宁 530023;
3 广西壮族自治区药用植物园/广西药用资源保护与遗传改良重点实验室, 广西 南宁 530023, correspAuthors=潘丽梅, authorNote=刘焕英: 刘焕英(1996-),硕士研究生,主要从事中药学研究。E-mail:17852186681@163.com, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=SRieesWTaewCYVPq+lMFuQ==, pdfFileSize=2176554, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=中央引导地方科技发展资金项目 (桂科ZY24212031); 广西自然科学基金项目 (2026GXNSFBA00640057,2023GXNSFAA026330); 广西岐黄学者培养项目 (GXQH202402))}, authors=[Author(id=1307432390087962668, tenantId=1146029695717560320, journalId=null, articleId=1304414961585901858, orderNo=null, firstName=null, middleName=null, lastName=null, nameCn=null, orcid=null, stid=null, country=null, authorPic=null, dead=null, email=null, emailSecond=null, emailThird=null, correspondingAuthor=null, authorType=null, ext={EN=AuthorExt(id=null, tenantId=null, journalId=1302319053441957962, articleId=1304414961585901858, authorId=1307432390087962668, language=EN, stringName=LIU Huanying, LEI Ming, XIAO Xiao, LI Linxuan, LI Cui, HUANG Yuan, HUO Juan, WAN Lingyun, PAN Limei, ZHANG Zhanjiang, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=null, address=null, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null), CN=AuthorExt(id=null, tenantId=null, journalId=1302319053441957962, articleId=1304414961585901858, authorId=1307432390087962668, language=CN, stringName=刘焕英, 雷明, 肖潇, 李林轩, 李翠, 黄媛, 霍娟, 万凌云, 潘丽梅, 张占江, firstName=null, middleName=null, lastName=null, prefix=null, suffix=null, authorComment=null, nameInitials=null, affiliation=null, department=null, xref=null, address=null, bio=null, bioImg=null, bioContent=null, aboutCorrespAuthor=null)}, companyList=null)], keywords=[Keyword(id=1304414962038886693, tenantId=1146029695717560320, journalId=1302319053441957962, articleId=1304414961585901858, language=CN, orderNo=1, keyword=积雪草), Keyword(id=1304414962110189862, tenantId=1146029695717560320, journalId=1302319053441957962, articleId=1304414961585901858, language=CN, orderNo=2, keyword=CaCYP2基因), Keyword(id=1304414962173104423, tenantId=1146029695717560320, journalId=1302319053441957962, articleId=1304414961585901858, language=CN, orderNo=3, keyword=基因克隆), Keyword(id=1304414962244407592, tenantId=1146029695717560320, journalId=1302319053441957962, articleId=1304414961585901858, language=CN, orderNo=4, keyword=原核表达), Keyword(id=1304414962307322153, tenantId=1146029695717560320, journalId=1302319053441957962, articleId=1304414961585901858, language=CN, orderNo=5, keyword=亚细胞定位), Keyword(id=1304414962445734186, tenantId=1146029695717560320, journalId=1302319053441957962, articleId=1304414961585901858, language=EN, orderNo=1, keyword=Centella asiatica (L.) Urban.), Keyword(id=1304414963691442475, tenantId=1146029695717560320, journalId=1302319053441957962, articleId=1304414961585901858, language=EN, orderNo=2, keyword=CaCYP2), Keyword(id=1304414963792105772, tenantId=1146029695717560320, journalId=1302319053441957962, articleId=1304414961585901858, language=EN, orderNo=3, keyword=gene cloning), Keyword(id=1304414963892769069, tenantId=1146029695717560320, journalId=1302319053441957962, articleId=1304414961585901858, language=EN, orderNo=4, keyword=prokaryotic expression), Keyword(id=1304414963964072238, tenantId=1146029695717560320, journalId=1302319053441957962, articleId=1304414961585901858, language=EN, orderNo=5, keyword=subcellular localization)], refs=null, funds=null, companyList=null, figs=null, attaches=null, journal=Journal(id=1302309778002903112, delFlag=0, nameCn=中草药, nameEn=Chinese Traditional and Herbal Drugs, nameHistory1=null, nameHistory2=null, issn=0253-2670, eissn=null, cn=12-1108/R, coden=null, periodic=3, 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=cGpSKCP11AF8PAOcTXYWfg==, journalPrice=null, startedYear=null, abbrevIsoEn=Chinese Traditional and Herbal Drugs, journalRemark=null, publicationField=null, createdTime=1788424446827, updatedTime=1788949289390, createdBy=18614031015, updatedBy=13041195026, firstLetterCn=Z, firstLetterEn=Z, subjectCode=Medical and Pharmaceutical Sciences, subjectName=null, subjectCodeEn=Medical and Pharmaceutical Sciences, subjectNameEn=null, picCn=cGpSKCP11AF8PAOcTXYWfg==, picEn=Xw//kxUC3ON4eHxev0QLhQ==, jcr=null, cjcr=null, exts=[JournalExt(id=1304511127375863983, language=CN, name=中草药, 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=1788949289411, updatedTime=1788949289411, createdBy=13041195026, updatedBy=13041195026, submissionGuidelinesUrl=, submissionAuthorUrl=https://www.tiprpress.com/zcy/author/login, submissionEditorUrl=https://www.tiprpress.com/zcy/editor/login, submissionReviewUrl=https://www.tiprpress.com/zcy/reviewer/login, submissionCeEditorUrl=, submissionAeEditorUrl=, option={"copyright":""}), JournalExt(id=1304511127442972848, language=EN, name=Chinese Traditional and Herbal Drugs, 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=1788949289427, updatedTime=1788949289427, createdBy=13041195026, updatedBy=13041195026, submissionGuidelinesUrl=, submissionAuthorUrl=https://www.tiprpress.com/zcy/author/login, submissionEditorUrl=https://www.tiprpress.com/zcy/editor/login, submissionReviewUrl=https://www.tiprpress.com/zcy/reviewer/login, submissionCeEditorUrl=, submissionAeEditorUrl=, option={"copyright":""})], databaseList=null, tenantJournalId=1302319053441957962, websiteList=[Website(id=1302319176408912052, webName=null, webTitle=null, webDomain=null, webCopyrigh=null, webIpcNo=null, seoTitle=null, seoKeywords=null, seoDescription=null, tenantJournalId=null, journalId=1302319053441957962, 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/zcy/CN, language=CN, createTime=1788426687576, createBy=18614031015, updateTime=1788427346252, updateBy=18614031015, name=中草药-中文, tplId=1146099689490845704, title=中草药, delFlag=0, indexPage=/home, props=[WebsiteProps(id=1302322043651904087, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1302319176408912052, code=articleTextType, value=kx, createTime=1788427371180, updateTime=1788427371180, creator=18614031015, updator=18614031015), WebsiteProps(id=1302322043593183828, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1302319176408912052, code=banner, value=null, createTime=1788427371166, updateTime=1788427371166, creator=18614031015, updator=18614031015), WebsiteProps(id=1302322043672875610, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1302319176408912052, code=grayFlag, value=0, createTime=1788427371185, updateTime=1788427371185, creator=18614031015, updator=18614031015), WebsiteProps(id=1302322043584795219, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1302319176408912052, code=logo, value=https://castjournals.cast.org.cn/joweb/zcy/CN/file/pic?fileId=uiD1gpiRqR++OLOz4iKzDg==, createTime=1788427371164, updateTime=1788427371164, creator=18614031015, updator=18614031015), WebsiteProps(id=1302322043689652828, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1302319176408912052, code=minRunFlag, value=0, createTime=1788427371189, updateTime=1788427371189, creator=18614031015, updator=18614031015), WebsiteProps(id=1302322043643515478, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1302319176408912052, code=picServerUrl, value=https://castjournals.cast.org.cn/joweb/zcy/CN/file/pic, createTime=1788427371178, updateTime=1788427371178, creator=18614031015, updator=18614031015), WebsiteProps(id=1302322043681264219, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1302319176408912052, code=silenceFlag, value=0, createTime=1788427371187, updateTime=1788427371187, creator=18614031015, updator=18614031015), WebsiteProps(id=1302322043601572437, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1302319176408912052, code=staticResourcePath, value=https://castjournals.cast.org.cn/joweb/cast_kjdb_cn_619/, createTime=1788427371168, updateTime=1788427371168, creator=18614031015, updator=18614031015), WebsiteProps(id=1302322043660292696, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1302319176408912052, code=themeColor, value=null, createTime=1788427371182, updateTime=1788427371182, creator=18614031015, updator=18614031015), WebsiteProps(id=1302322043668681305, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1302319176408912052, code=themeStyle, value=null, createTime=1788427371184, updateTime=1788427371184, creator=18614031015, updator=18614031015)]), Website(id=1302319176715096246, webName=null, webTitle=null, webDomain=null, webCopyrigh=null, webIpcNo=null, seoTitle=null, seoKeywords=null, seoDescription=null, tenantJournalId=null, journalId=1302319053441957962, 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/zcy/EN, language=EN, createTime=1788426687649, createBy=18614031015, updateTime=1788427341161, updateBy=18614031015, name=中草药-英文, tplId=1146101810881728533, title=Chinese Traditional and Herbal Drugs, delFlag=0, indexPage=/home, props=[WebsiteProps(id=1302322015206134340, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1302319176715096246, code=articleTextType, value=kx, createTime=1788427364398, updateTime=1788427364398, creator=18614031015, updator=18614031015), WebsiteProps(id=1302322015185162817, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1302319176715096246, code=banner, value=null, createTime=1788427364393, updateTime=1788427364393, creator=18614031015, updator=18614031015), WebsiteProps(id=1302322015227105863, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1302319176715096246, code=grayFlag, value=0, createTime=1788427364403, updateTime=1788427364403, creator=18614031015, updator=18614031015), WebsiteProps(id=1302322015176774208, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1302319176715096246, code=logo, value=https://castjournals.cast.org.cn/joweb/zcy/EN/file/pic?fileId=uiD1gpiRqR++OLOz4iKzDg==, createTime=1788427364391, updateTime=1788427364391, creator=18614031015, updator=18614031015), WebsiteProps(id=1302322015239688777, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1302319176715096246, code=minRunFlag, value=0, createTime=1788427364406, updateTime=1788427364406, creator=18614031015, updator=18614031015), WebsiteProps(id=1302322015201940035, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1302319176715096246, code=picServerUrl, value=https://castjournals.cast.org.cn/joweb/zcy/EN/file/pic, createTime=1788427364397, updateTime=1788427364397, creator=18614031015, updator=18614031015), WebsiteProps(id=1302322015235494472, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1302319176715096246, code=silenceFlag, value=0, createTime=1788427364405, updateTime=1788427364405, creator=18614031015, updator=18614031015), WebsiteProps(id=1302322015193551426, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1302319176715096246, code=staticResourcePath, value=https://castjournals.cast.org.cn/joweb/cast_kjdb_en_623/, createTime=1788427364395, updateTime=1788427364395, creator=18614031015, updator=18614031015), WebsiteProps(id=1302322015214522949, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1302319176715096246, code=themeColor, value=null, createTime=1788427364400, updateTime=1788427364400, creator=18614031015, updator=18614031015), WebsiteProps(id=1302322015218717254, tenantId=1146029695717560320, journalId=null, journalGroupId=null, siteId=1302319176715096246, code=themeStyle, value=null, createTime=1788427364401, updateTime=1788427364401, creator=18614031015, updator=18614031015)])], journalTitle=中草药, weixinUrl=null, journalUrl=https://www.tiprpress.com/zcy, iacademicId=null, status=1, seqNo=null, journalTitleEn=Chinese Traditional and Herbal Drugs, journalPhotoCn=cGpSKCP11AF8PAOcTXYWfg==, journalPhotoEn=Xw//kxUC3ON4eHxev0QLhQ==, journalFirstLetter=Z, 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, interPubPlatform=, interPubPlatformUrl=null), detailUrlCn=https://castjournals.cast.org.cn/joweb/zcy/CN/10.7501/j.issn.0253-2670.2026.07.023, detailUrlEn=https://castjournals.cast.org.cn/joweb/zcy/EN/10.7501/j.issn.0253-2670.2026.07.023, pdfUrlCn=https://castjournals.cast.org.cn/joweb/zcy/CN/PDF/10.7501/j.issn.0253-2670.2026.07.023, pdfUrlEn=https://castjournals.cast.org.cn/joweb/zcy/EN/PDF/10.7501/j.issn.0253-2670.2026.07.023, aliStartDate=null, aliEndDate=null, collectionFlag=false, citedCount=null, citedUrl=null, previewStatus=0, delFlag=0, hasFullText=0, orderTime=1788926361700, fullTextJson=null, articleText=null, reference=中国药典[S].一部. 2025:336.
黄晓玲,陈文,王湘,等.黎药积雪草植物药用成分研究进展[J].内江科技, 2021, 42(3):112-113.
秦慧真,林思,邓玲玉,等.积雪草苷的药理作用及机制研究进展[J].中国药房, 2021, 32(21):2683-2688.
古宝浚,史俊豪,刘晓芳,等.积雪草化学成分、药理作用研究进展及质量标志物预测分析[J].辽宁中医药大学学报, 2025, 27(7):35-43.
王迎迎,顾永政.积雪草苷对代谢相关脂肪性肝病大鼠Nrf2、ROS通路表达的影响[J].现代药物与临床,2024, 39(4):810-815.
Diniz L R L, Calado L L, Duarte A B S, et al. Centella asiatica and its metabolite asiatic acid:Wound healing effects and therapeutic potential[J]. Metabolites, 2023,13(2):276.
林琛,王斌,刘巨钊,等.积雪草化学成分和药理作用研究进展及其质量标志物预测分析[J].中医药信息,2023, 40(8):70-77.
魏艺聪,陈建雄,黄泽豪,等.基于matK序列的积雪草与其易混品的分子鉴定方法分析[J].世界中医药,2016, 11(5):900-902.
曹柯科,刘智谋,张胜,等.低共熔溶剂提高积雪草有效成分提取效率的研究[J].湖南林业科技, 2023,50(1):10-14.
Khalili H M, Mirjalili M H, Mohajeri M. Boosted centellosides production in Gotu kola(Centella asiatica)transgenic hairy roots elicited by gold and zinc nanoparticles[J]. Ind Crops Prod, 2025, 227:120796.
曹策,杨丽丽,宋鉴书,等.积雪草苷调控PIK3C3/DYNC2H1/TUB减轻转化生长因子-β1诱导的心肌成纤维细胞活化[J].中草药, 2025, 56(14):5108-5118.
Kim O T, Um Y, Jin M L, et al. A novel multifunctional C-23 oxidase, CYP714E19, is involved in asiaticoside biosynthesis[J]. Plant Cell Physiol, 2018, 59(6):1200-1213.
朱灵英,郭娟,张爱丽,等.参与植物三萜生物合成的细胞色素P450酶研究进展[J].中草药, 2019, 50(22):5597-5610.
田荣,谷巍,韦陈彬,等.植物三萜类成分生物合成中氧鲨烯环化酶与细胞色素P450的研究进展[J].南京中医药大学学报, 2021, 37(2):303-310.
杨成佳,成旭,胡冰,等. P450酶在植物三萜化合物生物合成中的催化与调控[J].生物加工过程, 2023,21(1):39-49.
覃雅,雷明,李林轩,等.积雪草CaPDS基因克隆、表达模式分析及过表达载体构建[J].中药材, 2025,48(8):58-78.
张顺仓,刘肖宇,邱琳,等.丹参SmCYP76S7基因的克隆、亚细胞定位和表达分析[J].中草药, 2024,55(13):4500-4507.
温丽翠.亚洲璃眼蜱源CYP450基因克隆表达及其功能分析[D].乌鲁木齐:新疆农业大学, 2024.
刘越圳,聂小山,尚进,等.茅苍术细胞分裂素羟化酶基因(AlCYP735A)的克隆与表达分析[J].中草药,2025, 56(9):3253-3262.
Yu H W, Wang H X, Liang X, et al. Telomere-to-telomere gap-free genome assembly provides genetic insight into the triterpenoid saponins biosynthesis in Platycodon grandiflorus[J]. Hortic Res, 2025, 12(5):uhaf030.
Zhang S X, Meng F Q, Pan X, et al. Chromosome-level genome assembly of Prunella vulgaris L. provides insights into pentacyclic triterpenoid biosynthesis[J]. Plant J,2024, 118(3):731-752.
Miettinen K, Pollier J, Buyst D, et al. The ancient CYP716family is a major contributor to the diversification of eudicot triterpenoid biosynthesis[J]. Nat Commun, 2017, 8:14153.
Song W L, Chen B Z, Feng L, et al. Telomere-to-telomere genome assembly and 3D chromatin architecture of Centella asiatica insight into evolution and genetic basis of triterpenoid saponin biosynthesis[J]. Hortic Res, 2025,12(5):uhaf037.
Park J W, Reed J R, Backes W L. The localization of cytochrome P450s CYP1A1 and CYP1A2 into different lipid microdomains is governed by their N-terminal and internal protein regions[J]. J Biol Chem, 2015, 290(49):29449-29460.
Lu H P, Edwards M, Wang Q Z, et al. Expression of cytochrome P450 CYP81A6 in rice:Tissue specificity,protein subcellular localization, and response to herbicide application[J]. J Zhejiang Univ Sci B, 2015, 16(2):113-122.
Wang R B, Zhang L Y, Li X, et al. Single-cell sequencing and mass spectrometry imaging reveal the multicellular compartmentalization map of plant triterpenes:A ginsenoside in Panax ginseng example[J]. Plant Biotechnol J, 2025, 23(10):4461-4476.
Yang L F, Yang Z, Liu M, et al. Integrated single-cell transcriptomics and spatial metabolomics unveil cellular differentiation and ginsenosides biosynthesis in Panax root tips[J]. Hortic Res, 2025, 12(11):uhaf202.
李娇珍,孙正海,蔡起航,等.基于LC-MS积雪草不同部位中化学成分分析[J].西部林业科学, 2022,51(4):75-79.
Sheibani N. Prokaryotic gene fusion expression systems and their use in structural and functional studies of proteins[J]. Prep Biochem Biotechnol, 1999, 29(1):77-90.
Porowińska D, Wujak M, Roszek K, et al. Prokaryotic expression systems[J]. Postepy Hig Med Dosw, 2013, 67:119-129.
李亚伟,刘增才,孙婷婷,等.暴马桑黄细胞色素P450家族CYP5150AW6基因克隆及表达分析[J].中草药,2022, 53(11):3448-3455.
赵桂红,张妮妮,高帅帅,等.菘蓝CYP83A1基因的克隆、表达特性及原核表达分析[J].基因组学与应用生物学, 2018, 37(1):302-312.
何听雨,逄雨,张远洋,等.高产乳酰-N-三糖Ⅱ大肠杆菌菌株的构建[J].生物技术通报, 2025, 41(11):143-152.
周金龙.针对常见融合标签的结合蛋白从头设计与亲和力表征[D].武汉:华中科技大学, 2024.
周小洁,邱星辉,曾晓芃.细胞色素P450原核功能表达系统概述[J].毒理学杂志, 2021, 35(3):241-245.
王世山,陈艳可,杨俊.膜蛋白原核表达的基因序列优化[J].生物技术通报, 2015, 31(12):50-55.
孟凡琦.北马兜铃阿朴啡类生物碱骨架形成相关CYP80的鉴定及功能分析[D].北京:北京协和医学院, 2024.
李可鑫.白桦CYP450基因的表达特性及在三萜合成中的功能鉴定[D].哈尔滨:东北林业大学, 2022.)