猪萨佩罗病毒YC2011株1D基因的克隆及原核表达  

Cloning and Prokaryotic Expression 1D Gene of YC2011 Strain of Porcine Sapelovirus

作　　者：陈俊伟[1] 张祥斌[2] 张云静[3] 周庆丰[1] 宋延华[1] 李薇[1] 陈峰[2] 薛春宜[3] 毕英佐[2] 曹永长[3] 

机构地区：[1]广东温氏食品集团股份有限公司,广东新兴527400 [2]华南农业大学动物科学学院,广东广州510642 [3]中山大学生命科学学院,广东广州510006

出　　处：《动物医学进展》2014年第1期68-72,共5页Progress In Veterinary Medicine

摘　　要：为研发猪萨佩罗病毒(PSV)检测试剂,根据PSV YC2011毒株核苷酸序列,设计针对1D基因的特异引物,以YC2011毒株为模板,利用RT-PCR方法,成功扩增出1D基因,将该基因与原核表达载体pET-32a(+)连接,转化表达菌株BL21。经PCR和测序鉴定,阳性菌株IPTG诱导表达,融合蛋白1D进行SDS-PAGE和Western blot检测分析。结果表明,成功构建了原核表达菌株,其所表达的融合蛋白分子质量约为51ku,且可被猪萨佩罗病毒阳性血清所识别。According to the porcine Sapelovirus YC2011 strain nucleotide sequence reported in the GenBank, we designed a pair of primers to amplify 1D gene of YC2011strain. The 1D gene was cloned into prokaryot- ic expression vector pET-32a（-b） and transformed into E. coli strain BL21. In order to obtain 1D protein, the positive strains were induced by IPTG, and then the expressed products were analyzed by SDS-PAGE and Western blot. The results of SDS-PAGE and Western blot demonstrated that the 1D gene of YC2011 strain was successfully expressed. The expressed 1D protein has 51 ku,and can be recognized by the posi- tive serum of Porcine sapelovirus.

关 键 词：猪萨佩罗病毒 1D基因 原核表达 

分 类 号：S852.659.6[农业科学—基础兽医学] Q786[农业科学—兽医学]