红汁乳菇中一个聚酮合酶基因的鉴定和表达分析  

Identification and Optimal Expression Medium of LhPKS1 Gene from Lactarius hatsudake

作　　者：原晓龙 罗婷 王毅 李云琴 杨文忠 YUAN Xiao-long;LUO Ting;Wang Yi;LI Yun-qin;YANG Wen-zhong(Yunnan Key Lab.of Forest Plant Cultivat′n&Exploit′n/Conserv.of Rare,Endangered&Endemic Forest Plants,Public Key Lab.of Ntl.Forest.&Grassland Administ′n,Yunnan Prov.Acad.of Forest.&Grassland,Kunming 650201)

机构地区：[1]云南省林业和草原科学院云南省森林植物培育与开发利用重点实验室/国家林业和草原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650201

出　　处：《微生物学杂志》2020年第5期18-25,共8页Journal of Microbiology

基　　金：国家自然科学基金项目(31860177);云南省林业科学院创新基金项目(MS2019-09)。

摘　　要：聚酮化合物具有丰富的生物活性,为了解红汁乳菇(Lactarius hatsudake)中聚酮合酶基因,从红汁乳菇基因组中分离并克隆得到LhPKS1基因,通过生物信息学分析推测其功能,并通过RT-PCR验证该基因的表达量。结果显示,LhPKS1基因全长cDNA含有6036 bp,编码2011个氨基酸残基,结构域顺序依次为SAT-KS-AT-PT-ACP-TE,该蛋白无跨膜结构和信号肽,聚类分析显示LhPKS1蛋白与参与生物合成苔色酸的真菌PKS蛋白聚为一支。在以10%肌醇、2%和10%的山梨醇为碳源添加物及以番茄浸粉为氮源添加物时,该基因表达量较高。研究有助于通过LhPKS1基因的过表达及异源表达,为大量获取苔色酸类化合物及其骨架提供参考。Polyketides have abundant bioactivities.In order to understand polyketide synthase gene(PKS),LhPKS1 gene was isolated and cloned from the genome of Lactarius hatsudake in this study,its function was predicted by the bioinformatics method,and its expression levels in medium including different carbon or nitrogen additives was detected.The results showed that the full-length of LhPKS1 gene has 6036 bp,and encodes 2011 amino acid residues;the sequence of structural domain of LhPKS1 protein successively was SAT-KS-AT-PT-ACP-TE;the protein had no transmembrane structure and signal peptide;the cluster analysis showed that the LhPKS1 protein and other fungus PKS protein that participated in the biosynthesis of orsellinic acid clustered into one branch;the gene expression of LhPKS1 was higher when making use of 10%inositol,2%and 10%sorbitol as carbon additives and tomato extract powder as nitrogen resource additives.This study was conducive to contribute some material to get a mass amount of orsellinic acids and their framework adopting over-expression and heterologous expression of LhPKS1 gene.

关 键 词：红汁乳菇(Lactarius hatsudake) 聚酮合酶 生物信息学分析 克隆 表达 

分 类 号：Q93-331[生物学—微生物学]