尼帕病毒N基因的原核表达  

Prokaryotic expression of Nipah virus N gene

作　　者：梁栋[1,2] 涂长春[3] 石清明[2] 张超雄[2] 陈锚锚[2] 邓成玉[2] 范泉水[2] 

机构地区：[1]重庆第三军医大学流行病学教研室,400038 [2]成都军区疾病预防控制中心 [3]军事医学科学院军事兽医研究所

出　　处：《西南国防医药》2015年第3期255-258,共4页Medical Journal of National Defending Forces in Southwest China

摘　　要：目的利用尼帕病毒(NiV)的N基因进行原核表达,探讨NiV N蛋白作为检测NiV存在的抗原的可能性。方法利用pET-28a(+)作为表达载体,构建pET-NiV N重组质粒,通过IPTG诱导使其表达蛋白,并用Western blot方法进行原核表达NiV N蛋白的检测。结果通过PCR、酶切和连接反应,成功构建N蛋白的pET-NiV N重组质粒,大小为1596 bp;利用IPTG诱导pET-NiV N重组质粒在宿主菌中表达N蛋白,SDS-PAGE结果显示,55 k D有特异蛋白条带,与蛋白预计大小相符,经Bandscan软件扫描分析,重组蛋白量占菌体总蛋白量的12%;Western blot法检测显示,55 k D的条带显色良好,证明是本实验所需要得到的目的蛋白。结论使用原核表达系统成功表达了NiV的N蛋白,为其进一步的抗体制备、活性鉴定及研究应用奠定了基础。Objective To explore the possibility of Nipah virus （ NiV） N protein as an antigen to detect the presence of NiV by using the prokaryotic expression of NiV N gene .Methods pET-28a （＋） was used as an expression vector in order to construct the recombinant plasmid of pET-NivN, which was induced by IPTG to express the protein; and then, Western blot method was used to detect the prokaryotic expression of NivN protein .Results By means of PCR , enzyme digestion and ligation reaction , the recombinant plasmid of pET-NivN was constructed successfully , with a size of 1596 bp;IPTG was used to induce the recombinant plasmid of pET-NivN to express N protein in the host bacteria .SDS-PAGE results showed that 55 kD has a specific protein band with the expected protein size; according to the Bandscan software scanning and analysis , the amount of recombinant protein was 12% of the total bacteria protein .Western blot assay showed that the band of 55 kD has a good color , and this proved that it was just the protein to be obtained in the experiment .Conclusion The prokaryotic expression system can express N protein of NiV successfully , and this lays solid foundation for further antibody preparation , activity evaluation , and research application .

关 键 词：尼帕病毒 N基因 原核表达 

分 类 号：R373[医药卫生—病原生物学]