Expression of soluble H1N1 haemegglutinin in two distinct insect baculovirus expression systems
10.3760/cma.j.issn.0254-5101.2011.04.015
- VernacularTitle:利用两种真核昆虫表达系统表达可溶性甲型H1N1血凝素蛋白
- Author:
Hao WANG
;
Chi MA
;
Lianxian CUI
;
Denian BA
;
Wei HE
- Publication Type:Journal Article
- Keywords:
Influenza virus haemegglutinin;
Baeularvirus expression system;
BaculoGold system;
Bac-to-Bac system;
Haemegglutinination test
- From:
Chinese Journal of Microbiology and Immunology
2011;31(4):345-349
- CountryChina
- Language:Chinese
-
Abstract:
Objective To express functional haemegglutinin(HA)protein in two different bacularvirus expression systems.Methods The whole open reading frame of A/Sichuan/1/2009(H1N1)HA was obtained by synthesis,and the HA protein were expressed in insect cells by two different bacularvius expression systems:BaculoGold system and Bac-to-Bac system. Soluble HA protein was identified by Western blot and haemegglutination test. Results The correct full length of HA gene was obtained and cloned into pAcGP67B and pFAST Bacl vectors,respectively.After 3 rounds of virus amplifyjng by re-infection of Sf9 cells,the HA protein was detected in supematant of BaculoGold system and in intracellular of Bac-to-Bac system which is much better than the former.Purified HA protein was positive not only identified by Western blot,but also detected by haemegglutinin test. Conclusion Functional HA protein was successfully expressed in two distinct bacularvirus expression systems,of which the Bac-to-Bac bacularvirus expression system is more suitable for expression of A/Sichuan/1/2009(H1N1)HA protein.
