牛疙瘩皮肤病病毒ORF132基因的克隆及其原核表达  

Cloning and prokaryotic expression of ORF132 of Lumpy skin disease virus

作　　者：周雪梅[1,2] 聂福平[2,3] 王昱[2,3] 肖进文[2,3] 聂奎[1] 李应国[2,3] 

机构地区：[1]西南大学动物科技学院,重庆北碚400715 [2]重庆出入境检验检疫局,重庆江北400020 [3]重庆进出口食品安全工程中心,重庆江北400020

出　　处：《中国预防兽医学报》2012年第4期320-322,共3页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家质检总局科研项目(2010IK0015);国家质检总局科研项目(2009IK027)

摘　　要：为原核表达牛疙瘩皮肤病病毒(LDSV)ORF132并对该基因进行生物信息学分析,本研究采用PCR扩增LSDV的ORF132基因,将其亚克隆至pGEX-6p-1载体中,构建重组质粒pGEX-LSDV-ORF132。生物信息学分析表明,LSDV的ORF132与Neethling vaccine LW1959株LW132、NI-2490株LSDV132和NW-LW株LD132的相应推导氨基酸同源性分别为100%、100%和94.4%,与山羊痘病毒、绵羊痘病毒相应推导氨基酸的同源性为20.7%～36%。SDS-PAGE和western blot分析表明,pGEX-LSDV-ORF132在大肠杆菌BL21中经IPTG诱导主要以包涵体形式存在,其重组蛋白大小约为48.95 ku,并且与特异性抗体具有抗原反应活性。In response to the risk of Lumpy skin disease virus（LSDV） spreading to China,it is necessary to establish inspection methods for LSDV infection.Through bioinformatics analysis,it was suggested that LSDV-ORF132 shared high homology with LW132（Neethling vaccine）,LSDV132（NI-2490） and LW132（NW-LW）,the amino acid homologies were 100%,100% and 94%,respectively.In comparison with corresponding amino acid sequences of the original gene in capripox virus,the homologies were ranged from 20.7% to 36%.In this experiment,a 542 bp fragment of LSDV-ORF132 was amplified by PCR and inserted into prokaryotic expression vector pGEX-6p-1 for expressed in E.coli.The recombinant protein was 48.95 ku,and specifically reacted with LSDV antibody,indicating the recombinant protein possessed antigenic activity.

关 键 词：牛疙瘩皮肤病病毒 LSDV-ORF132基因 序列分析 原核表达 

分 类 号：S852.65[农业科学—基础兽医学]