首页|Lithocarols类化合物生物合成基因litI的表达及其启动子功能分析

Lithocarols类化合物生物合成基因litI的表达及其启动子功能分析

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  • 国家科技期刊平台
  • NETL
  • NSTL
  • 万方数据
[目的]Lithocarols为新型的多异戊烯基二苯甲酮类化合物,具有良好的抗肿瘤活性.对lithocarols生物合成基因litI进行克隆和表达纯化,并对该基因的启动子进行功能鉴定,为lithocarols的生物合成及转录调控奠定分子生物学基础.[方法]将litI基因扩增后进行原核表达,采用镍亲和层析初步纯化目的蛋白LitI,利用生物信息学方法分析该蛋白的性质和结构.同时扩增litI基因启动子片段,构建荧光素酶表达系统,分析启动子的转录活性,利用PlantCARE启动子分析网站对litI基因启动子的功能组件进行预测.[结果]LitI蛋白为亲水性蛋白,其相对分子量为 51 kD,二级结构包括 51.32%的α-螺旋、8.99%的延伸链、3.95%的β-转角以及35.75%无规则卷曲.litI基因启动子具有较强的转录活性且在大肠杆菌中具有启动氨苄青霉素抗性基因表达的功能,其功能组件包含TATA box和CAAT box.[结论]通过异源表达获得了LitI蛋白,分析了其性质和结构,并鉴定了具有较强转录活性的litI基因启动子片段.
Expression of Lithocarols Biosynthesis Gene litI and Functional Analysis of Its Promoter
[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.

Phomopsis lithocarpuspolyisoprenyl benzophenonesbiosynthetic genepromoterheterologous expressionbioinformatics

李梦然、叶伟、李赛妮、张维阳、李建军、章卫民

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佛山科学技术学院生命科学与工程学院,佛山 528225

广东省科学院微生物研究所华南应用微生物国家重点实验室广东省菌种保藏与应用重点实验室,广州 510070

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

国家自然科学基金国家自然科学基金广东省基础与应用基础研究基金广东省科学院专项

32370081322000392020A15151104972021GDASYL-20210103021

2024

10.13560/j.cnki.biotech.bull.1985.2023-1225

生物技术通报
中国农业科学院农业信息研究所

生物技术通报

CSTPCD北大核心
影响因子：0.505
ISSN：1002-5464
年,卷(期)：2024.40(6)
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