堆型艾美耳球虫早熟系与母株11种抗原基因表达水平的比较分析  

Comparative analysis of the expression levels of 11 antigenic genes between Eimeria acervulina vaccine and parent strains

作　　者：邹尚书 梁建英[1] 王新秋[1] 何媛[1] 程田[1] 翁亚彪[1] 林瑞庆[1] 

机构地区：[1]华南农业大学兽医学院,广东广州510642

出　　处：《中国兽医科学》2014年第1期33-38,共6页Chinese Veterinary Science

基　　金：广东省科技计划项目(2012A020100001);家畜疫病病原生物学国家重点实验室开放课题(SKLVEB 2012KFKT003)

摘　　要：应用荧光定量qRT-PCR技术比较堆型艾美耳球虫早熟系及其母株孢子化卵囊的11种抗原基因的表达情况。提取堆型艾美耳球虫早熟系及其母株孢子化卵囊的总RNA,反转录成cDNA,在GenBank中查找堆型艾美耳球虫11种抗原基因并设计相应引物,进行荧光定量qRT-PCR扩增,用熔解曲线、扩增曲线和循环阈值(Ct值)的变化等比较分析堆型艾美耳球虫早熟系及其母株的不同基因的扩增特异性和相对表达量。结果显示,11种抗原基因得到表达;与疫苗母株相比,早熟致弱的疫苗株大部分抗原基因的表达量下降,其中HSP70基因表达量下降明显。结果表明,早熟系毒力的减弱可能与某些抗原基因表达量下降有关。The purpose of the present study was to assay and compare the expression levels of 11 anti- genic genes in the sporulated oocysts of Eimeria acervulina precocious line and its parent strain using quantitative RT-PCR. Total RNA of the sporulated oocysts of these two parasite samples was extracted^nd reversely transcribed into eDNA respectively. The primers for the 11 antigenic genes experimentally evalu- ated previously were designed based on the corresponding sequences available in GenBank and applied to fluorescent quantitative RT-PCR reactions. The dissociation curves, amplification curves and cycle thresh- old（Ct） were used as parameters for evaluating these genes＇ relative expression levels of differences be- tween the two parasite samples. In result, the expression levels of most of these 11 antigenic genes in the precocious line decreased,especially HSP70 gene. The result showed that the virulence attenuation in the precocious line might be due to the declined expression of these antigenic genes.

关 键 词：堆型艾美耳球虫 抗原基因 表达水平 荧光定量PCR 

分 类 号：S852.723[农业科学—基础兽医学]