摘要
牛樟芝(Antrodia camphorata)是一种珍稀食药用菌,能产生具有抗癌活性的倍半萜类化合物。对牛樟芝基因组进行分析,获得倍半萜合酶基因序列并设计特异引物,提取在Glu培养基(麦芽浸粉6 g/L,酵母提取物3 g/L,葡萄糖40 g/L)上生长的牛樟芝菌丝体的RNA,利用RT-PCR技术克隆得到倍半萜合酶基因AcTPS1。AcTPS1基因c DNA全长为969 bp,编码323个氨基酸,根据系统进化树可知AcTPS1氨基酸序列与其他9种真菌倍半萜合酶聚为一类。AcTPS1拥有典型倍半萜合酶的结构域(RRSRSATAEAYACFIW),之后检测AcTPS1在不同培养基上的菌丝中的表达结果显示不同碳源中,只有葡萄糖作为碳源时该基因表达,不同氮源中,以番茄浸粉和酪蛋白胨为氮源时该基因表达。说明AcTPS1是一类诱导型表达的基因。为利用发酵培养以及异源表达手段获得牛樟芝活性化合物提供参考。
Antrodia camphorata was rare edible and medicinal fungi and it can produce sesquiterpene compounds with anti-cancer activity.In this research,A.camphorata of genome was analyzed,then sesquiterpene synthase gene sequence was obtained and primers were designed according to this sequence.The RNA of mycelium on the Glu-medium was extracted.The sesquiterpene synthase gene of AcTPS1was cloned by RT-PCR which full-length of cDNA was969bp,this sequence can encode323amino acids.According to phylogenetic tree,the amino acid sequence of AcTPS1was clustered with other9sesquiterpene synthases which has a typical sesquiterpene synthase domain(RRSRSATAEAYACFIW).The expression level of AcTPS1in mycelium on different media was shown,gene was expressed only when glucose was used as the carbon source among the different carbon sources,the gene also expressed when tomato powder and casein peptone as the nitrogen source in different nitrogen sources.Indicating that AcTPS1is a class of inducible expression of genes.This research make an important foundation for obtaining the active compounds by using fermentation method and heterologous expression.
作者
赵能
陈中华
原晓龙
王娟
杨宇明
王毅
Zhao Neng;Chen Zhonghua;Yuan Xiaolong;Wang Juan;Yang Yuming;Wang Yi(The Key Laboratory of Forest Plants Cultivation and Utilization, Yunnan Academy of Forestry, Kunming 650204, China;The Key Laboratory of Rare and Endangered Forest Plants of State Forestry Administration, Yunnan Academy of Forestry, Kunming 650204, China;College of Forestry, Southwest Forestry University, Kunming 650204, China)
出处
《微生物学杂志》
CAS
CSCD
2017年第5期13-19,共7页
Journal of Microbiology
基金
云南省对外科技合作计划项目(2015IA004)
国家自然科学青年科学基金项目(31400488)
云南省面上项目(2016FB055)
云南省重点实验室开放基金项目(ZZCX2016-11)
关键词
牛樟芝
倍半萜合酶
基因克隆
基因表达
Antrodia camphorata
sesquiterpene synthase
gene cloning
gene expression
