烟夜蛾谷胱甘肽S-转移酶基因的克隆、序列分析与表达  

Molecular cloning,sequence analysis and expression of a glutathione S-transferase gene from Helicoverpa assulta (Guenée) (Lepidoptera:Noctuidae)

作　　者：杨新影[1] 李亮[1] 安世恒[1] 罗梅浩[1] 原国辉[1] 郭线茹[1] 

机构地区：[1]河南农业大学植物保护学院,郑州450002

出　　处：《昆虫学报》2011年第6期648-656,共9页Acta Entomologica Sinica

基　　金：河南省杰出青年科学基金项目(074100510013)

摘　　要：为探明谷胱甘肽S-转移酶(GSTs)在昆虫嗅觉识别中的作用,本研究采用RT-PCR和RACE方法,从烟夜蛾Helicoverpa assulta(Guenée)雄虫触角中克隆获得了1个GSTs基因的全长cDNA序列(GenBank登录号为EU289223)。将该基因推导的氨基酸序列与其他物种的GSTs进行同源性比对和系统发育分析,发现该蛋白属于昆虫特异性Epsilon家族成员,因此将该基因命名为HaGSTe1。同时从烟夜蛾基因组DNA中克隆获得了该基因序列,发现序列中含有5个内含子,长度分别为415,513,296,333和269bp。利用半定量RT-PCR和实时荧光定量PCR方法对HaGSTe1在雌、雄虫不同组织的表达进行了定性和定量分析,结果显示,该基因在雌、雄虫的头部(去掉触角和喙)、触角、喙、胸、足、翅以及雌虫的腹部均有表达,并且在雄虫触角中的表达量最高,且显著高于雌虫触角,这种表达情况提示其可能与触角中性信息素及其他外源物质的分解有关。In order to explore the function of glutathione S-transferases （GSTs） in olfactory recognition of insects,the full-length cDNA of a novel glutathione S-transferase gene was cloned from the antennae of Helicoverpa assulta by using RT-PCR and RACE methods （GenBank accession no.EU289223）.Based on the sequence identity and the phylogenetic tree of the amino acids of GSTs in H.assulta and other species,the sequence obtained was found to belong to the Epsilon family,hence named as HaGSTe1.Genomic structure analysis of HaGSTe1 revealed that HaGSTe1 contained five introns with the length of 415,513,296,333 and 269 bp,respectively.The qualitative and quantitative analyses of the expression of HaGSTe1 in male and female moths were conducted using semi-quantitative RT-PCR and real-time PCR.The results showed that HaGSTe1 transcript was clearly detected in the head （with antennae and proboscis removed）,antenna,proboscis,thorax,legs,and wings of both male and female and abdomen of the female.HaGSTe1 transcript was significantly more highly expressed in male antennae than in female antennae,suggesting that it may be involved with the decomposition of pheromones and other xenobiotics in the antenna.

关 键 词：烟夜蛾 谷胱甘肽S-转移酶 基因克隆 半定量RT-PCR 实时荧光定量PCR 

分 类 号：Q966[生物学—昆虫学]