云南农业大学学报2012，Vol.27Issue(4)：530-534,5.DOI:10.3969/j.issn.1004-390X(n).2012.04.012
猪瘟病毒Npro基因的克隆表达
Molecular Cloning and Expression of Npro Gene of the Classical Swine Fever Virus
摘要
Abstract
In order to select an eukaryotic expression vector for Npro protein, and to study the relationship between Npro and major histocompatibility complex ( MHC) at different time point, a full length Npro gene of classical swine fever virus ( CSFV) of Shimen was amplified and cloned by RT-PCR. The results showed that the Npro gene contained 592 bp. The recombinant plasmid and expression vector pcDNA3. 0 were digested by the restriction enzymes Bam H I and Xho I , after the amplified fragments was subcloned into the expression vector pcDNA3. 0, the recombinant plasmid P-Npro was obtained. The insert position, size and reading frame were determined by PCR, restriction digestion and the sequence analysis. The recombinant P-Npro plasmid expressed Npro was used to transfect PK-15 cells in vitro by liposome infection protocol. The results showed that after 24 h and 48 h posttransfeclion, Npro protein was detected by Western-blot analysis with anti-His monoclonal antibody, and it was found that the Npro protein was located.关键词
猪瘟病毒/Npro基因/真核表达Key words
Classical swine fever virus/ Npro gene/ eukaryotic expression分类
农业科技引用本文复制引用
高斌,冯励,李永强,付晓萍..猪瘟病毒Npro基因的克隆表达[J].云南农业大学学报,2012,27(4):530-534,5.基金项目
云南省教育厅科学研究基金项目(2011Y443). （2011Y443）
