Co-expressing M1 and NA genes of infuenza H1N1 virus in Baculovirus expression system
10.3760/cma.j.issn.1003-9279.2015.01.010
- VernacularTitle:H1N1亚型流感病毒M1和NA基因在昆虫杆状病毒系统中的共表达
- Author:
Aijun CHEN
1
;
Lihong YAO
;
Pengwei XU
;
Zhiqing ZHANG
;
Jianqiang GUO
Author Information
1. 中国疾病预防控制中心病毒病预防控制所 卫生部医学病毒和病毒病重点实验室
- Keywords:
Influenza virus;
Proteins;
Neuraminidase;
Baculovirus
- From:
Chinese Journal of Experimental and Clinical Virology
2015;29(1):29-31
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the recombinant baculovirus co-expressing M1 and]NA genes of influenza H1N1 virus.Methods The M1 and NA genes of influenza virus (A/PR8/8/34) were amplified by PCR and then inserted into the pFastBacdual (pFBD)donor plasmid to construct the recombinant pFBDM1-NA.pFBD-M1-NA was transformed into DH10Bac E.coli competent cells containing bacmid vector to construct the recombinant shuttle vector rBacmid-M1-NA.Sf9 cells were transfected with rBacmid-M1-NA and then the recombinant baculovirus rBac-M1-NA was obtained.The virus titer was tested by plaque formation,M1 and NA genes in genome were identified by PCR,and the expression of M1 and NA gene by IFA.Results PCR result proved that pFBD-M1-NA and rBacmid-M1-NA were constructed correctly.The recombinant baculovirus rBac-M1-NA titer was 1 × 107pfu/ml.IFA analysis showed that M1 and NA genes were expressed in sf9 cells infected with rBac-M1-NA.Conclusion The recombinant baculovirus coexpressing M1 gene and NA gene of infuenza H1 N1 virus was successfully established.
