A型流感病毒NP基因核酸疫苗的构建与体外表达鉴定

Evaluation on construction and expression in vitro of nucleic acid vaccine of nucleoprotein of influenza virus A

目的构建A型流感病毒核蛋白核酸疫苗,研究核酸疫苗主动免疫保护效应。方法从A型流感病毒逆转录获得NP基因,与表达载体pSECTAG2HygroA构建核酸疫苗,采用脂质体转染法转染VERO细胞,经ELISA鉴定,A型流感病毒NP基因核酸疫苗在VERO细胞中得到了表达。结果结果显示,在转染后36h得到最大化表达,其A450达到0.382。在36到60h时间范围内,NP蛋白表达量基本没有变化(在48h和60h的A450分别为0.385和0.387)。结论成功构建了A型流感病毒核蛋白核酸疫苗,为进一步研究该疫苗体内实验奠定了基础。

Objective\ Constructing nucleic acid vaccine of influenza virus A to study the protective effects. Methods NP gene was reverse transcripted from influenza virus A and linked with pSECTAG 2/Hygro A to constructed nucleic acid vaccine of influenza virus A. The constructed nucleic acid vaccine was transinfected into VERO cell resorting to lipofectamineTM2000. NP gene of influenza virus A was expressed in VERO cell by the method of ELISA. Results The results showed that the NP gene was expressed to the highest level at 36h after transinfection with the A_(450) value of 0.382. From 36 h to 60 h after transinfection, the expression of NP gene was stable ~the A_(450) value at 48h and 60h was 0.385 and 0.387, respectively). Conclusion The nucleic acid vaccine of influenza virus A was successfully constructed, which sets up groudworks for the research of effection of nucleic acid vaccine in vivo.