Comparison of three methods in construction fusion gene of influenza A virus Nucleoprotein
10.3760/cma.j.issn.1003-9279.2012.01.024
- VernacularTitle:构建甲型流感病毒核蛋白融合基因三种方法的比较研究
- Author:
Bo LIU
1
;
Bao-Lin WANG
;
Ling CHEN
;
Jing YAN
;
Jian-Yong ZHANG
;
Hong-Xuan HE
;
Hong ZHANG
Author Information
1. 遵义医学院附属医院
- Keywords:
Influenza A virus;
Nuclear proteins;
Green fluorescent protein;
Genes
- From:
Chinese Journal of Experimental and Clinical Virology
2012;26(1):70-74
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop a best method of constructing influenza NP fusion gene containing enhanced green fluorescent protein (EGFP).Methods The full-length NP gene of influenza A was amplified by RT-PCR and was inserted into an eukaryotic expression vector pEGFP-N1 in order to construct a fusion gene of pEGFP-N1-NP using three different methods.Method one,NP gene containing restriction endonucleases and pEGFP-N1 were both digested using the same restrict enzymes and ligated,yielding the fusion gene of pEGFP-N1-NP. Method two,NP gene was cloned into pMD19-T Vector to construct a plasmid of pMD19-T-NP.The pMD19-T-NP cloned into pEGFP-N1 to construct the fusion gene of pEGFP-N1-NP; Method three,NP gene containing restriction endonucleases was cloned into pMD19-T Simple Vector to construct a plasmid of pMD19-T-NP.The pMD19-T-NP cloned into pEGFP-N1 to construct the fusion gene of pEGFP-N1-NP.Results The fusion gene of recombinant eukaryotic expression vector pEGFP-N1-NP was successfully constructed by using method three.Conclusions The full-length NP gene is obtained and its fusion gene of recombinant eukaryotic expression plasmid is successfully constructed.This study provides foundation for further understanding the biological function of NP protein and the mechanism of diseases induced by influenza A virus.
