艾纳香BbCHS基因的克隆及其原核表达载体的构建

Gene Cloning of BbCHS in Blumea balsamifera DC.and Its Construction of Prokaryotic Expression Vector

查尔酮合酶(chalcone synthase,CHS)是艾纳香(Blumea balsamifera DC.)黄酮类化合物代谢途径的关键酶之一。本研究以产自贵州省罗甸县道地艾纳香为研究材料,参照艾纳香叶的转录组数据,设计PCR引物,成功克隆得到查尔酮合成酶基因的完整cDNA序列,利用生物信息学分析软件对其进行相关分析,包括氨基酸性质、信号肽、跨膜域、多序列比对、进化树构建以及蛋白的二、三级结构预测等,并且构建了CHS原核表达载体,在BL21宿主菌中可以表达目的蛋白,为黔产艾纳香中CHS的生物学研究提供依据,同时为艾纳香黄酮类物质的生物合成提供理论指导。

Chalcone synthase(CHS)is one of the key enzymes during the flavonoids metabolic pathway in Blumea balsamifera DC..Taken Blumea balsamifera DC.as research materials,this study refered the transcriptome results,designed the PCR primers,cloned the full c DNA sequence of CHS and analyzed its biological information,including characteristics of amino acids,signal peptide,trans-membrane domain,multi-sequence blast,construction of phylogenetic tree,secondary and tertiary structure of CHS protein.And prokaryotic expression vector of CHS that can express target proteins in BL21 bacterial was constructed.This will lay a foundation for the biological study of CHS and provide theoretical guidance for the biosynthesis of flavonoids in Blumea balsamifera DC..