猪细小病毒NS1基因真核表达载体pEGFP-N1-NS1的构建与表达  

Construction and expression of eukaryotic expression vector pEGFP-N1-NS1 for porcine parvovirus NS1 gene

作　　者：周雍[1] 张驰[1] 王朋涛 韩丽[2] 靳晓慧[2] 魏战勇[1,2] ZHOU Yong;ZHANG Chi;WANG Pengtao;HAN Li;JIN Xiaohui;WEI Zhanyong(Engineering College of Animal Husbandry and Veterinary Science,Henan Agricultural University,Zhengzhou 450002,China;Key Laboratory for Animal-derived Food Safety of Henan Province,Zhengzhou 450002,China)

机构地区：[1]河南农业大学牧医工程学院,河南郑州450002 [2]河南省动物性食品安全重点实验室,河南郑州450002

出　　处：《河南农业大学学报》2018年第3期356-360,共5页Journal of Henan Agricultural University

基　　金：国家自然科学基金项目(U140410985);河南省高校科技创新人才项目(15HASTIT028)

摘　　要：根据Gen Bank上发表的PPV的基因组序列,设计扩增非结构蛋白NS1基因的特异性引物。通过PCR方法克隆NS1基因,经双酶切及与pEGFP-N1载体连接构建了真核表达载体pEGFP-N1-NS1,将双酶切和测序鉴定正确的质粒转染至293T细胞,经过荧光观察和PCR方法鉴定。结果表明,克隆了猪细小病毒NS1基因的全序列1989 bp,成功构建了NS1基因的真核表达载体,并在293T细胞中转染表达。According to the sequences of PPV genes in the Gen Bank,the special primers for nonstructural protein NS1 gene were designed. The NS1 gene of PPV virus was amplified by PCR and cloned into pEGFP-N1 vector to construct a plasmid,named pEGFP-N1-NS1. The plasmids were identified by double enzyme digestion and sequencing. The correcting plasmids were transfected into293 T cells and identified by fluorescence and PCR methods. The results showed that the recombinant eukaryotic expression vector p EGFP-N1-NS1 was successfully constructed; and the NS1 protein was finally expressed in 293 T cells.

关 键 词：猪细小病毒 真核载体 NS1 

分 类 号：S828[农业科学—畜牧学]