Expression of the hemagglutinin and neuramidinase gene of influenza A virus H1N1 in Pichia methanolica.
- Author:
Ye ZHANG
1
;
Zaijiang YU
;
Li XIN
;
Yongkun CHEN
;
Qihui TANG
;
Yubao CHEN
;
Qingxuan CHEN
;
Yuelong SHU
Author Information
1. Department of Influenza, Nattonat Institute of Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
- Publication Type:Journal Article
- MeSH:
Cloning, Molecular;
Genetic Vectors;
genetics;
Hemagglutinins;
biosynthesis;
genetics;
Influenza A Virus, H1N1 Subtype;
genetics;
Neuraminidase;
biosynthesis;
genetics;
Pichia;
genetics;
metabolism;
Recombinant Proteins;
biosynthesis;
genetics;
immunology
- From:
Chinese Journal of Biotechnology
2010;26(8):1068-1073
- CountryChina
- Language:Chinese
-
Abstract:
On the basis of successful cloning the full length hemagglulinin (HA) and neuramidinase (NA) gene and sequence analysis of influenza virus H1N1, part of the gene was ligated into pMETA. Expression vectors pMETA/HA (52-1 557 bp) and pMETA/NA (121-1 263 bp) were constructed and expressed in pMAD16 induced by methanol. Recombinant protein was purified through Ni2+ affinity chromatography. Western blotting and ELISA were used to determine the antigenic activity of the recombinant protein. SDS-PAGE showed that the recombinant capsid gene could be overexpressed in Pichia methanolica. ELISA and Western blotting showed that the recombinant protein had antigenicity.
