新城疫病毒xx08毒株血凝素-神经氨酸酶基因主要抗原区原核表达及鉴定

摘要

In this study,the HNii75-K367 gene fragment of HN gene epitope concentration region of Newcastle disease virus (NDV) was cloned into pET-32a( + ) vector to construct the expression plasmid pET32-HNI175-K367. The plasmid was transformed into E. coli BL21 (ED3) and induced by IPTG for HNI175-K367 protein expression. Then,the expressed protein was separated by SDS-PAGE and identified by western blot assay. The results showed that the HNi175-k367 protein was overly expressed and the molecular weight was about 38 kD. The expressed protein specifically reacted with anti-NDV antiserum, indicating that the prokaryotically expressed target protein had good antigencity.%利用基因工程技术构建新城疫病毒HN基因抗原表位集中区HNI175-K367基因片段(523-1101位)的重组表达质粒pET32-HNI175-K367.将该质粒转化大肠杆菌BL21( ED3),经IPTG诱导,SDS-PAGE鉴定表明,HNI175-K367蛋白得到高效表达,其分子量约为38 kD.Western-blot分析证实,HNI175-K367蛋白与鸡新城疫病毒高免血清发生特异性反应,表明利用原核表达系统获得的重组蛋白具有良好的反应原性.