Establishment of mammalian cell lines for constitutive expression of influenza virus matrix protein 2.
- Author:
Ai-Jun CHEN
1
;
Jian-Qiang GUO
;
Li-Hong YAO
;
Ji-Yong YIN
;
Zhi-Qing ZHANG
Author Information
1. Molecular Virology and Genetic Engineering Laboratory, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China. chenajsun@yahoo.com.cn
- Publication Type:Journal Article
- MeSH:
Animals;
CHO Cells;
Cell Culture Techniques;
Cell Line;
Cricetinae;
Cricetulus;
Influenza A Virus, H2N2 Subtype;
chemistry;
Recombinant Proteins;
biosynthesis;
Viral Matrix Proteins;
biosynthesis;
genetics
- From:
Chinese Journal of Virology
2013;29(1):7-11
- CountryChina
- Language:Chinese
-
Abstract:
To establish a mammalian cell line for stable expression of the matrix protein 2 (M2) of influenza virus type A. M2 gene was amplified by PCR from the influenza virus strain A/PR/8/34. The PCR product was cloned into eukaryotic expression vector pcDNA5/FRT. After identification with restriction enzyme digestion, the plasmid was co-transfected with plasmid pOG44 which expressed Flp in Flp-In-CHO cells. The target gene was integrated into chromosome of CHO cells by homologous recombination in vivo. Recombinant CHO-M2 cell lines were selected for hygromycin B resistance. A total of 15 recombinant cell strains with high expression of M2 protein were screened by hygromycin, and the expression of M2 protein was determined by IFA and Western blot. After subculturing for 10 passages, the presence of M2 gene in the CHO-M2 cells was confirmed by PCR, and the expression of M2 protein were proved by IFA and Western blot. We successfully constructed a mammalian cell line which stably expressed M2 protein of influenza virus type A. The cell line will be useful for studies on function of M2 protein and provide tools for novel influenza virus vaccine development.
