鸡胚致死孤儿病毒Ⅲa蛋白基因的克隆、表达及其抗原性鉴定  

Cloning and Expression of Protein Ⅲa Gene of Chicken Embryo Lethal Orphan Virus(CELOV)and Identification of Its Antigenicity

作　　者：周斌[1] 齐香荣[1] 杨耀武[2] 芦银华[1] 陈溥言[1] 

机构地区：[1]南京农业大学农业部动物疫病诊断与免疫重点开放实验室,南京210095 [2]沈阳农业大学动物医学系,沈阳110161

出　　处：《农业生物技术学报》2003年第6期616-619,共4页Journal of Agricultural Biotechnology

摘　　要：通过随机引物 PCR 鉴定了鸡(Gallus domesticus)胚孤儿致死病毒(CELOV)污染鸭肝炎病毒鸡胚尿囊液种毒。根据GenBank 上已公布的核苷酸序列,自行设计一对引物。PCR 扩增到1 728 bp 的特异性条带,将其在大肠杆菌(Escherichia coli)中高效表达,产物蛋白经 Western blot 鉴定为阳性后,又长程免疫小白鼠获得抗Ⅲa蛋白的抗血清。间接免疫荧光实验(IFA)鉴定蛋白产物具有抗原性。It was identified that chicken(Gallus domesticus)embryo lethal virus(CELOV)polluted duck viral hepatitis seed virus of chicken embryonic allantochoroin liquid by PCR.In order to find out the harmfulness of this virus,a fragment 1728 bp was amplified by PCR from the genome of CELOV and cloned into a prokaryotic expression vector pET-32a(+)and expressed it in Escherichia coli efficiently,based on Ⅲa gene sequence data.The fused protein was proved positive by Western blot and used to immunize the mouse and positive antibody was obtained.And the antigenicity of protein Ⅲa was tested by IFA.

关 键 词：鸡胚致死孤儿病毒Ⅲa蛋白基因 克隆 表达 抗原性 

分 类 号：Q78[生物学—分子生物学] S831[农业科学—畜牧学]