伪狂犬病病毒糖蛋白G基因的结构分析及其原核表达  

Analysis of the Structure and Prokaryotic Expressing of the Glycoprotein G Gene of Pseudorabies Virus

作　　者：闵平[1] 张楚瑜[1] 潘兹书[1] 

机构地区：[1]武汉大学病毒学研究所,武汉430072

出　　处：《中国病毒学》2002年第2期166-171,共6页Virologica Sinica

摘　　要：利用PCR技术扩增了伪狂犬病毒湖北株 (PRVHB)糖蛋白G(gG)基因 ,进行了序列测定和分析。结果显示扩增和测序片段长 180 4bp ,G +C含量 6 8.78%。gG基因ORF长 15 0 0bp ,编码 5 0 0个氨基酸组成的多肽。与PRVRice株 gG基因比较 ,两者核苷酸及推导的氨基酸序列同源性分别为 98%、84.1%。 32 0～ 380位之间的氨基酸序列存在较大差异。根据序列分析结果 ,选取 gG基因长短不同的两个片段分别克隆到原核表达载体 pET2 8a(+)进行表达。经SDS PAGE和Dot ELISA分析证实 ,表达出分子量大小分别约为 5 5kD和 6 3kD的特异性gG多肽 ,这为深入阐明PRV gG基因结构与功能及研制 gGThe glycoprotein G ( gG ) gene of pseudorabies virus Hubei strain (PRV HB) was amplified by PCR, sequenced and analyzed. The results revealed that the fragment cloned and sequenced was 1804 base pairs(bp) with a G+C content of 68.78% and an ORF containing 1500bp to encode a protein of 500 amino acids.Comparison of the gG gene of PRV HB with PRV Rice strain showed the sequence homologies of the nucleotide and deduced amino acid were 98% and 84.1%, respectively. Most of these differences were located within amino acid residues 320 to 380. According to the nucleotide sequence analysis, two different length segments of gG gene were cloned into the prokaryotic vector pET28a(+) and expressed. The specific polypeptides of gG , whose molecular weights were about 55kD, 63kD respectively, were identified by SDS-PAGE and Dot-ELISA.This results might be contribute to the study of the structure and function of gG gene and gG-ELISA diagnosis kit of PRV.

关 键 词：伪狂犬病病毒 糖蛋白G 原核表达 核苷酸序列 氨基酸 序列 基因表达 基因结构 

分 类 号：S852.65[农业科学—基础兽医学]