嗜虫书虱β3- actin基因克隆及定量PCR方法的建立

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 ento-mophila 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.