新城疫病毒HN基因功能结构域的原核表达

本研究对新城疫病毒(Newcastle disease virus,NDV)分离株sc05的HN基因进行了克隆、序列测定,在此基础上将其C端功能结构域76－571 aa片段亚克隆到pET32a(＋)表达载体上,构建重组表达质粒pET32a-HN,鉴定正确后转化进BL21 plysS(DE3)细胞,筛选出阳性克隆,用1 mmol/L IPTG 诱导表达,并对表达产物进行鉴定。SDS-PAGE电泳结果显示,HN基因功能结构域片段在 BL21 plysS(DE3)细胞中实现融合表达,表达产物大小约为76 ku,Western blotting试验证实其能与NDV阳性血清反应。本研究为进一步研究HN蛋白功能结构域的免疫原性和研制鸡新城疫HN基因工程疫苗奠定了基础。

Prokaryotic Expression of Function Domain of the Newcastle Disease Virus HN Gene

In this study,HN gene of Newcastle disease virus(NDV) isolate sc05 was cloned,sequenced,on this basis,the 76 to 571 aa fragment of C-terminal function domain was subcloned into pET32a(＋) expression vector,obtained recombinant expression plasmid pET32a-HN. pET32a-HN was identificated to be correct, and it was transfored into BL21 plysS(DE3) cells. Selected positive clones, induced expression by 1 mmol/L IPTG, and the expression products were identified. SDS-PAGE electrophoresis showed that HN gene function domain fragments in BL21 plysS(DE3) cells achieved fusion expression, size of the recombinant protein was about 76 ku, Western blotting confirmed the biological activity of recombinant protein. This work was benefit for further study of the functional domains of the HN protein immunogenicity and the development of the HN gene of Newcastle disease vaccine project.