猪瘟病毒E_2基因主要抗原区的克隆及原核表达  

Cloning of the Major Antigen Region of E_2 Gene of Hog Cholera Virus and Expression in Escherichia coli

作　　者：张永国 刘湘涛[1] 韩雪清[1] 刘希成[2] 张彦明[2] 谢庆阁[1] 

机构地区：[1]中国农业科学院兰州兽医研究所农业部畜禽病毒学重点实验室,兰州730046 [2]西北农林科技大学畜牧兽医学院,杨凌712100

出　　处：《生物工程学报》2002年第5期605-608,共4页Chinese Journal of Biotechnology

基　　金：国家重点基础研究发展规划 (G19990 119)~~

摘　　要：利用反转录PCR(RT PCR)和套式PCR(nestPolymeraseChainReaction ,nPCR)技术扩增出当前猪瘟流行毒(广西玉林株 ,GXYL)与中国猪瘟兔化弱毒 (C 株 )兔脾组织毒E2 基因的主要抗原区 ,将其克隆到表达载体pPROEX HTb中 ,获得重组质粒pPROEX GXYL和pPROEX C ,经PCR、酶切和序列分析鉴定表明 ,插入的位置、大小和读码框均正确。SDS PAGE检测表明 ,经重组质粒pPROEX GXYL和pPROEX C转化、诱导的受体菌能表达E2 基因主要抗原区蛋白 ,Western blot检测表明 。The major antigen region of E 2 gene of Hog Cholera Prevalent Strain (Guangxi Yuling Strain) and Chinese Hog Cholera Lapinised Virus (C-strain) derived from hog and rabbit spleen tissue, was amplified by reverse transcription polymerase chain reaction(RT-PCR) and the nested Polymerase Chain Reaction (nPCR).After the amplified fragments were cloned into the expression vector pP ROEX-HTb, the recombinant plasmids pP ROEX-GXYL and pP ROEX-C were obtained. The insert position ,the size and the reading frame were right by PCR,restriction digestion and the sequence analysis. SDS-PAGE indicated that both of the reciepient germs transducted and induced by the recombinant plasmids pP ROEX-GXYL and pP ROEX-C could express the major antigen region of E 2 gene. Western-blot indicated that the expressed antigen protein could be recognized by the positive serum of CSFV.

关 键 词：猪瘟病毒 E2基因 主要抗原区 克隆 原核表达 

分 类 号：Q786[生物学—分子生物学]