非洲猪瘟病毒p72蛋白杆状病毒表达系统的构建及其抗原性分析  

Expression and identification of African swine fever virus protein p72 expressed in insect cells

作　　者：侯绍华[1] 柏丽华[1] 郭晓宇[1] 贾红[1] 姜一曈[1] 朱鸿飞[1] 

机构地区：[1]中国农业科学院北京畜牧兽医研究所动物医学研究室,北京100193

出　　处：《中国预防兽医学报》2012年第1期33-36,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家十一五科技支撑计划项目(2009BADB4B00)

摘　　要：为研究真核表达非洲猪瘟病毒(ASFV)主要结构蛋白p72,本研究从含ASFV p72基因全序列的重组质粒pGEX-6p-p72中扩增出1 941 bp的p72全长基因,将其插入于pFastBac HTa杆状病毒载体中,构建了重组质粒pFastBac HTa-p72,转化至感受态细胞DH10Bac中,获得重组杆粒rBacmid-p72。再将其转染至sf9昆虫细胞,获得重组杆状病毒。Western blot和夹心法ELISA分析表明ASFV p72基因在昆虫细胞sf9中获得了正确表达,重组p72蛋白可以被特异性抗ASFV血清、p72单克隆抗体识别,表明该蛋白特异性强、活性稳定,具有良好的抗原性。为进一步研究p72蛋白的结构、功能和免疫学特性奠定基础。To express nucleocapsid p72 protein of African swine fever virus（ASFV）,the p72 gene was amplified by PCR and cloned into pFastBac HTa vector.The resultant recombinant plasmid was transformed into DH10Bac competent cells to construct recombinant Bacmid-p72 which was transfected into Sf9 insect cells to generate recombinant baculovirus.The p27 protein was expressed in Sf9 cells and identified by western blot and sandwich ELISA.The recombinant p72 protein was recognized by anti-ASFV positive serum and p72 MAb,which could be used for ASFV detection method development.

关 键 词：非洲猪瘟病毒 p72蛋白 杆状病毒表达 抗原性 

分 类 号：S852.65[农业科学—基础兽医学]