嗜虫书虱β-actin基因克隆及定量PCR方法的建立  

Cloning of β-actin Gene and Establishment of Quantitative Real-time PCR from Liposcelis Entomophila

作　　者：唐培安[1] 王进军[2] 宋伟[1] 

机构地区：[1]南京财经大学食品科学与工程学院,南京210003 [2]西南大学重庆市昆虫学及害虫控制工程重点实验室,重庆400716

出　　处：《中国粮油学报》2011年第11期71-75,共5页Journal of the Chinese Cereals and Oils Association

基　　金：国家自然科学基金(31000828)

摘　　要：旨在建立嗜虫书虱的实时荧光定量PCR分析方法,为嗜虫书虱基因的定量分析提供技术支持。利用RT-PCR方法,从嗜虫书虱体内克隆获得了β-actin基因cDNA片段(GenBank登录号:FJ041117),该基因片段长度为822 bp,编码273个氨基酸残基(从第3个碱基开始编码)。根据此β-actin基因的序列设计引物,建立了基于SYBR Green I染料技术的实时荧光定量PCR方法。建立的嗜虫书虱β-actin实时荧光定量PCR法扩增效率高、检测范围广、检测周期短,为β-actin作为内参基因进行嗜虫书虱功能基因的定量分析奠定了基础。The purpose of this study was to establish a quantitative real-time PCR method for Liposcelis entomophila and provide useful methodological basis for quantitative analysis of L.entomophila genes.The sequence of the β-actin gene cDNA fragment of Liposcelis entomophila was amplified by reverse transcript polymerase chain reaction（RT-PCR）,which contained 822 base pairs（bp） and encoded 273 amino acid（GenBank accession number： FJ041117）.According to the sequence of β-actin gene above,a quantitative real-time PCR method based on SYBR Green I dye was developed.The results showed that the quantitative real-time PCR method established in the study had the advantages of high efficiency,wide linear range and less time.The results of this study might provide a basis for β-actin used as a reference gene to analyze the Liposcelis entomophila gene quantitatively.

关 键 词：嗜虫书虱 Β-ACTIN基因 实时荧光定量 PCR内参基因 

分 类 号：Q786[生物学—分子生物学]