一个新型牛樟芝PR-PKS基因的克隆及表达分析  

Cloning and Peptidomic Analysis of A New PR-PKS Gene in Taiwanofungus camphoratus

作　　者：原晓龙[1] 华梅[1] 陈剑[1] 王娟[1] 杨宇明[1] 王毅[1] YUAN Xiao-long;HUA Mei;CHEN Jian;WANG Juan;YANG Yu-ming;WANG Yi(Yunnan Prov. Acad. of Forestry,Yunnan Prov. Key Lab. of Cultiv′n & Exploit′n of Forest Plants,Conserv′n of Rare,Endangered & Endemic Forest Plants,Public Key Lab. of the State Forestry Administ′n,Kunming 650204)

机构地区：[1]云南省林业科学院云南省森林植物培育与开发利用重点实验室国家林业局云南珍稀濒特森林植物保护和繁育重点实验室,云南昆明650204

出　　处：《微生物学杂志》2019年第1期11-19,共9页Journal of Microbiology

基　　金：国家自然科学地区基金项目(31860177);云南省面上基金项目(2016FB055);云南省对外科技合作计划项目(2015IA004)

摘　　要：为了解牛樟芝中聚酮化合物的生物合成机理及聚酮合酶基因功能,从牛樟芝基因组挖掘并克隆得到一个部分还原型PKS(PR-PKS)基因(AcPKS3),并对其进行生物信息学分析及表达谱分析。结果显示,AcPKS3(GenBank登录号:MG988206)DNA全长8 286 bp,有22个内含子,其外显子共编码2 285个氨基酸;结构域依次为KS-AT-KR-ACP-SDR,各结构域的活性保守位点为β-酮基合成酶(DTACSS)、酰基转移酶(GHSAGETA)、酮基还原酶(YLLVGGIG)、酰基转移酶(YGLDSITSA)、短链醇脱氢酶与NAD(P)H结合的N端保守序列(ITGTTGSFG)及活性保守位点(YTESK);AcPKS3与6-甲基水杨酸合成酶的亲缘关系较近;不同碳源中葡萄糖,不同氮源中牛肉浸粉、酪蛋白胨、土豆蛋白胨可促进AcPKS3基因表达。本研究为牛樟芝聚酮合酶功能研究及牛樟芝基因资源利用提供参考。In order to acquaint mechanism of biosynthesis of polyketides and the function of polyketides synthase ( PKS ) gene in Taiwanofungus camphoratus, the present study digged the PKS gene from T. camphoratus genome, cloned and obtained partial reduction type PKS (PR-PKS) gene (AcPKS3), and carried out its bioinformatic analysis and expression spectrum. The results showed that AcPKS3 (GenBank ID: MG988206) had 8 286 bp having 22 introns, 23 extrons, and its extrons encoded 2 285 amino acids;the structural domain successively was KS-AT-KR-ACP-SDR;and the conservative active sites of every structural domains were β-ketosynthase (DTACSS), acyl transferase (GHSAGETA), keto-reductase (YLLVGGIG), acyl transferase (YGLDSITSA), short chain alcohol dehydrogenase of NAD(P)H-binding with N terminal conservative sequence (ITGTTGSFG) as well as active conservative site (YTESK);the AcPKS3 was fairly close to 6-methylsalicyclic acid synthase (6MSAS) consanguineously;glucose in the different carbon source, beef extract powder in different nitrogen source, casein peptone and potato peptone could promote the expression of AcPKS3 gene. This study had laid foundation for further exploration of T. camphoratus AcPKS3 function and the application of T. camphoratus gene resource.

关 键 词：牛樟芝 部分还原型PKS 生物信息学分析 基因表达 

分 类 号：Q786[生物学—分子生物学]