中国畜牧兽医2013，Vol.40Issue(2)：18-22,5.
新城疫病毒核衣壳蛋白基因在大肠埃希菌中的表达、纯化及其活性分析
Prokaryotic Expression and Purification of NDV NP Gene in Escherichia coli and its Activity Analysis
摘要
Abstract
The nucleocapsid protein (NP) gene of Newcastle disease virus (NDV) was cloned into a prokaryotic expression vector, and then using E. coli BL2KDE3) to express this protein. NP gene was amplified from cRNA which was reverse transcription of RNA of NDV La Sota strain, then the gene was cloned into pET-30a. PCR,digestion and sequencing were used to identify positive plasmid, which named pET-NDV NP; the identified recombined plasmid was expressed by E. coli BL2(DE3) and induced by IPTG, the expressed products were purified by Ni-NTA Ni+ affinity column; SDS-PAGE was used to analyze the expressed protein; Western blotting was used to identify the reaction activity. The results showed that the sequence of NP gene was consistent with predicted size, 1470 bp. PCR, digestion and sequencing results showed that the recombinant plasmid pET-NDV NP was correct. NP protein was successfully expressed by E. coli BL2KDE3), then the recombinant protein was purified by Ni-NTA Ni+ affinity column and the concentration of the purified protein was 3 mg/mL. The recombinant protein could react with the antibody of NDV NP protein using the method of Western blotting assay. In summary, the recombinant plasmid pET-NDV NP containing NP gene was constructed and recombinant protein was successfully expressed in E. coli BL21 (DE3) and purified.关键词
新城疫病毒/NP蛋白/原核表达/纯化Key words
Newcastle disease virus (NDV)/NP protein/ prokaryotic expression purification分类
生物科学引用本文复制引用
徐彦召,王青,杭柏林,尚田田,赵志雨,胡建和..新城疫病毒核衣壳蛋白基因在大肠埃希菌中的表达、纯化及其活性分析[J].中国畜牧兽医,2013,40(2):18-22,5.基金项目
河南省高等学校青年骨干教师资助计划(2005461). （2005461）
