番鸭细小病毒 NS1基因的克隆与原核表达

摘要

本研究利用 PCR 方法扩增番鸭细小病毒（Muscovy duck parvovirus，MPV）非结构蛋白 NS1基因，并将其克隆至原核表达载体 pET32a (+)，构建重组表达质粒 pET32a-NS1。将其转化受体菌 E.coli Rosetta 感受态细胞，经 IPTG 诱导表达。SDS-PAGE 和 Western blot 试验结果表明，NS1基因在大肠杆菌中得到了表达，并且可与抗 MPV 的多克隆抗体发生特异性反应。MPV NS1基因的成功表达为研制 MPV 的诊断试剂盒、基因工程疫苗等奠定了基础。%The NS1 gene of Muscovy duck parvovirus (MPV) was amplified by PCR and inserted into multiple clone sites of pET32a ( + ) vector. The insertion of NS1 gene was confirmed in restriction endonuclease analysis and sequencing. The recombinant plasmid was transformed into E. coli Rosetta and NS1protein was expressed with induction of 1. 0 mmol/ L IPTG. The recombinant NS1 protein was visualized in SDS-PAGE and recognized by MPV specific antibody in Western blot. The successful expression of NS1 gene laid the foundation for development of diagnosis reagent kit and genetically engineering vaccine.