牛樟芝1个AcHMGS基因的克隆与表达分析  

Cloning and Expression Analysis of a HMGS Gene from Antrodia comphorata

作　　者：罗娅娜 郑元 原晓龙 王毅 LUO Ya-na;ZHENG Yuan;YUAN Xiao-long;WANG Yi(College of Forestry,Southwest Forestry University,Kunming Yunnan 650224,P.R.China;Yunnan Academy of Forestry and Grassland Sciences,Kunming Yunnan 650201,P.R.China)

机构地区：[1]西南林业大学林学院,云南昆明650224 [2]云南省林业与草原科学院,云南昆明650201

出　　处：《西部林业科学》2020年第4期176-181,共6页Journal of West China Forestry Science

基　　金：国家自然科学基金项目(31860177);云南省博士后科研基金;云南省博士后定向培养资助。

摘　　要：为了探明牛樟芝三萜化合物合成途径中的关键限速酶羟甲基戊二酰辅酶A合成酶的调控机制,为其三萜生物合成酶表达机制的研究提供参考,从牛樟芝中分离并克隆出1个基因,将其命名为AcHMGS,利用生物信息学分析软件了解其结构特性,分析该基因在不同碳氮源添加物培养基上的表达情况。结果表明,AcHMGS基因含有完整的开放阅读框,全长为1440bp,编码574个氨基酸;含有6个外显子、5个内含子。由蛋白质活性保守位点分析可知,AcHMGS蛋白包含由21个氨基酸组成的活性中心"GNTDIEGVDSKNACYGSTASL"和5个参与底物催化的保守氨基酸,其中2个能影响酶的活性。分子系统进化分析显示,AcHMGS蛋白与根纤维孔菌蛋白具有较高的同源性。AcHMGS基因的表达量在不同碳氮源添加物的培养基上差异显著,以甘露醇为碳源添加物、牛肉浸粉为氮源添加物的诱导表达量最高。In order to provide references for the study of triterpene synthase expression mechanism.We research a key rate-limiting enzyme in the pathway of synthesis triterpene in Antrodia comphorata termed Hydroxymethylglutaryl-CoA synthase.We cloned a gene named AcHMGS from Antrodia comphorata to understand it’s structural properties by bioinformatics software.Analyzed the expression of the gene in medium with different carbon and nitrogen sources.The results showed that the AcHMGS gene contained a complete open reading frame(OFR)with length of 1440 bp,encoding 574 amino acids.It contains 6 exons and 5 introns.According to the analysis of conservative sites of protein activity,the AcHMGS contain the active center"GNTDIEGVDSKNACYGSTASL",which composed of Twenty-one amino acids and five conserved amino acids involved in substrate catalysis,two of them can affect the activity of the enzyme.Molecular phylogenetic analysis showed that proteins of AcHMGS was homologous with Fibroporia radiculosa proteins.The expression level of AcHMGS gene was significantly different in the medium with different carbon and nitrogen source additives.The expression level was the highest with mannitol as carbon source additive and beef extract as nitrogen source additive.

关 键 词：牛樟芝 基因克隆 羟甲基戊二酰辅酶A合成酶 诱导表达 

分 类 号：S567.3[农业科学—中草药栽培] Q786[农业科学—作物学]