Article(id=1322943712967402494, tenantId=1146029695717560320, journalId=1322258771877134531, issueId=1322942934613635295, articleNumber=null, orderNo=null, doi=10.14067/j.cnki.1673-923x.2026.05.016, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1748361600000, receivedDateStr=2025-05-28, revisedDate=1753545600000, revisedDateStr=2025-07-27, acceptedDate=null, acceptedDateStr=null, onlineDate=1793343960276, onlineDateStr=2026-10-30, pubDate=1779638400000, pubDateStr=2026-05-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1793343960276, onlineIssueDateStr=2026-10-30, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1793343960276, creator=13701087609, updateTime=1793343960276, updator=13701087609, issue=Issue{id=1322942934613635295, tenantId=1146029695717560320, journalId=1322258771877134531, year='2026', volume='46', issue='5', pageStart='1', pageEnd='222', issueExtLink='null', onlineDate='null', pubDate='1779638400000', pubDateStr='2026-05-25', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1793343774703, creator='13701087609', updateTime=1793344699149, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1322946812499329386, tenantId=1146029695717560320, journalId=1322258771877134531, issueId=1322942934613635295, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1322946812499329387, tenantId=1146029695717560320, journalId=1322258771877134531, issueId=1322942934613635295, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=158, endPage=166, ext={EN=ArticleExt(id=1322943713135174655, articleId=1322943712967402494, tenantId=1146029695717560320, journalId=1322258771877134531, language=EN, title=Cloning and eukaryotic expression analysis of
CYP6620B1 gene of cytochrome P450 family in
Sanghuangporus baumii, columnId=null, journalTitle=Journal of Central South University of Forestry & Technology, columnName=null, runingTitle=null, highlight=null, articleAbstract=
【Objective】 To clone the full-length cDNA of CYP6620B1 gene, which is a key gene in the downstream pathway of triterpenoid synthesis in S. baumii and perform bioinformatics analysis and eukaryotic expression analysis of the gene, with a view to providing a theoretical basis for the elucidation of the mechanism of triterpenoid biosynthesis pathway of S. baumii and the enhancement of triterpenoid production.
【Method】 The transcript level of CYP6620B1 gene was detected by qRT-PCR at different developmental stages of the mycelium, protocorm and zygote of S. baumii. The full-length cDNA sequence of the target gene was obtained by PCR amplification, and the physicochemical properties and spatial structure of the protein were analyzed by bioinformatics online software. The recombinant Saccharomyces cerevisiae strain INVSc/pYES2-CYP6620B1 was obtained by LiAC/PEG method, and the total triterpene content of it was detected by microplate reader.
【Result】 The transcript level of CYP6620B1 gene in different developmental stages of S. baumii showed a trend of increasing and then decreasing, which coincided with the reported trend of total triterpenoids in S. baumii, suggesting that it might be involved in the regulation of triterpenoid synthesis, and the full-length cDNA of CYP6620B1 was cloned and obtained as 1 629 bp, encoding 542 amino acids, without signal peptide and transmembrane structure, and the sub The results of phylogenetic tree showed that the CYP6620B1 protein of Taiwanofungus camphoratus had the highest homology with that of S. baumii, and the molecular docking results showed that the CYP6620B1 protein had the lowest binding energy and good docking conformation with lanosterol; the CYP6620B1 gene was successfully expressed in the Saccharomyces cerevisiae strain INVSc, and the total triterpene content of recombinant Saccharomyces cerevisiae strain INVSc/pYES2-CYP6620B1 was significantly higher than that of the control strain.
【Conclusion】 In this study, the CYP6620B1 gene, which may be involved in the triterpene synthesis pathway of S. baumii, was successfully cloned and analysed, which lays the foundation for an in-depth investigation of the gene's function in the triterpene biosynthesis pathway of S. baumii.
, authors=Wenqi XIAO, Yawei LI, Nannan GENG, Chenyuan SI, Li ZOU, authorsList=Wenqi XIAO, Yawei LI, Nannan GENG, Chenyuan SI, Li ZOU, authorCompany=null, correspAuthors=Li ZOU, authorNote=null, correspAuthorsNote=null, copyrightStatement=© 2026 Journal of Central South University of Forestry & Technology, copyrightOwner=Journal of Central South University of Forestry & Technology, 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=1322943713504273408, articleId=1322943712967402494, tenantId=1146029695717560320, journalId=1322258771877134531, language=CN, title=暴马桑黄细胞色素P450家族
CYP6620B1基因克隆及真核表达分析, columnId=1322924272984929210, journalTitle=中南林业科技大学学报, columnName=生物学, runingTitle=null, highlight=null, articleAbstract=
【目的】 克隆暴马桑黄三萜类化合物合成下游途径中关键CYP6620B1基因的全长cDNA,并对其进行生物信息学分析和真核表达分析,以期为阐明暴马桑黄三萜生物合成途径机制及提高三萜产量提供理论依据。
【方法】 采用qRT-PCR技术检测CYP6620B1基因在暴马桑黄菌丝体、原基、子实体等不同发育阶段的转录水平。采用PCR扩增技术获得目的基因的cDNA序列全长,采用生物信息学在线软件分析其蛋白的理化性质和空间结构。采用LiAC/PEG法获得重组酿酒酵母菌株INVSc/pYES2-CYP6620B1,并采用酶标仪法检测其总三萜含量。
【结果】 CYP6620B1基因在暴马桑黄不同发育阶段的转录水平呈现先上升再下降的趋势,与已报道的暴马桑黄中总三萜含量的变化趋势相吻合,提示其可能参与三萜合成调控,进而克隆获得了CYP6620B1基因cDNA全长,为1 629 bp,编码542个氨基酸,不具有信号肽和跨膜结构,亚细胞定位于细胞质和内质网;系统进化分析结果表明,暴马桑黄CYP6620B1蛋白和牛樟芝蛋白同源性最高;分子对接结果显示,CYP6620B1蛋白与羊毛甾醇的结合能最低并具有良好的对接构象;CYP6620B1基因成功在酿酒酵母菌株INVSc中实现表达,且重组酿酒酵母菌株INVSc/pYES2-CYP6620B1中总三萜含量显著高于对照菌株。
【结论】 成功克隆得到可能参与暴马桑黄三萜合成途径的CYP6620B1基因,并对其进行分析,为深入探究该基因在暴马桑黄三萜生物合成途径中的功能奠定了基础。
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