Construction of recombinant baculovirus co-expressing M1 and HA of influenza A virus.
- Author:
Peng-Wei XU
1
;
Jian-Qiang GUO
;
Li-Hong YAO
;
Ai-Jun CHEN
;
Xiao-Yu LIU
;
Xian-Yin ZENG
;
Zhi-Qing ZHANG
Author Information
1. College of Life Science and Science, Sichuan Agricultural University, Yaan 625014, China. mengzhonren@163.com
- Publication Type:Journal Article
- MeSH:
Animals;
Baculoviridae;
genetics;
metabolism;
Cell Line;
Cloning, Molecular;
Gene Expression;
Genetic Vectors;
genetics;
metabolism;
Hemagglutinin Glycoproteins, Influenza Virus;
genetics;
immunology;
Influenza A Virus, H1N1 Subtype;
genetics;
immunology;
Spodoptera;
Transfection;
Viral Matrix Proteins;
genetics;
immunology
- From:
Chinese Journal of Virology
2012;28(3):231-236
- CountryChina
- Language:Chinese
-
Abstract:
The M1 and HA genes of H1N1 influenza virus were amplified and then cloned into the pFastBac dual donor plasmid. The recombinant pFastBac Dual-M1-HA was identified by restriction enzyme digestion. After the pFastBacdual-M1-HA was transformed into the baculovirus shuttle plasmid (bacmid) in DH10Bac competent cells, the colonies were identified by antibiotics and blue-white selection. The rBac-mid-M1-HA was verified by PCR and transfected into S f9 cells to produce recombinant baculovirus (rBac-M1-HA). Gene insertion of rBac-M1-HA was verified and the expression of M1 and HA genes was analyzed by IFA and Western-blot, demonstrating M1 and HA were co-expressed successfully. This study provides the foundation for researching the formation mechanism of influenza VLP and developing new influenza vaccines.
