生物技术通报2024，Vol.40Issue(6)：310-318,9.DOI:10.13560/j.cnki.biotech.bull.1985.2023-1225
Lithocarols类化合物生物合成基因litI的表达及其启动子功能分析
Expression of Lithocarols Biosynthesis Gene litI and Functional Analysis of Its Promoter
摘要
Abstract
[Objective]Lithocarols are a novel class of polyisoprenyl diphenylketone compounds,which have a good antitumor activity.The lithocarols biosynthesis gene litI was cloned,expressed and purified,the promoter of the gene litI was functionally identified,which will lay a molecular foundation for the biosynthesis and transcriptional regulation of lithocarols.[Method]The biosynthetic gene litI was cloned and expressed in Escherichia coli,and the expressed protein LitI was preliminarily purified by nickel affinity chromatography column.The properties and structure of the protein were analyzed by bioinformatics method.The litI gene promoter fragment was amplified,the luciferase expression system was constructed to analyze the transcriptional activity of the promoter,and its functional components were predicted by the PlantCARE promoter analysis website.[Result]LitI protein is a hydrophilic protein,which has a relative molecular weight of 51 kD and a secondary structure including 51.32%α-helix,8.99%extended chain,3.95%β-angle,and 35.75%irregular curling.The gene litI promoter shows strong transcription activity,and it could initiate the transcription of ampicillin resistance gene in E.coli,and litI promoter contained TATA box and CAAT box.[Conclusion]LitI protein is obtained by heterologous expression,its properties and structure are analyzed,and the promoter fragment of litI gene with strong transcription activity is identified.关键词
Phomopsis lithocarpus/多异戊烯基二苯甲酮类化合物/生物合成基因/启动子/异源表达/生物信息学Key words
Phomopsis lithocarpus/polyisoprenyl benzophenones/biosynthetic gene/promoter/heterologous expression/bioinformatics引用本文复制引用
李梦然,叶伟,李赛妮,张维阳,李建军,章卫民..Lithocarols类化合物生物合成基因litI的表达及其启动子功能分析[J].生物技术通报,2024,40(6):310-318,9.基金项目
国家自然科学基金项目(32370081,32200039),广东省基础与应用基础研究基金项目(2020A1515110497),广东省科学院专项资金项目(2021GDASYL-20210103021) （32370081,32200039）
