截短NDVF蛋白的原核表达

Prokaryotic Expression of Truncate F Protein Gene of Newcastle Disease Virus

根据NDV LaSota株F基因已知的抗原表位,对F蛋白进行分段表达。应用RT-PCR方法分段扩增F基因,并将其克隆到pET30a(+)原核表达载体上,得到重组质粒pET30-F780和pET30-F760,将质粒导入BL21(ED3)感受态中,经IPTG诱导表达。表达的重组蛋白通过SDS-PAGE和Westem-blotting方法进行鉴定。表达的两段蛋白大小约为31.1 kDa和27.9 kDa,与预期的蛋白分子量大小相符。Western blot分析表明重组蛋白可以和NDV抗体发生特异性反应。成功构建了原核表达质粒pETF780和pET-F760,并获得了高效表达,通过Western blot分析表明重组蛋白具有良好的免疫反应性。

According to the epitopes of F gene of NDV LaSota strain, the F gene truncated two fragments were expressed in E.coli BL21 （ED3） strain. The two truncate F gene amplified by RT-PCR were inserted into pET30 （＋）, a prokaryotie expression vector. The recombinant plasmid pET30-FTso and pET30-F760 were transformed into BL21 （ED3） competent cells. SDS-PAGE and Western- blotting screened the recombinant proteins induced by IPTG in E.coli. The size of the recombinant proteins were 31.1 kDa and 27.9 kDa,which were also consistent with those expected. Western-blotting showed that FTso and F760 were of immunogenicity.The recombinant plasmids were constructed, called pET-FTso and pET-F760, which were expressed the corresponding proteins with better immunoreactivity.