Construction of recombinant baculovirus co-expressing M1 and HA of influenza A virus

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 c...

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Published inBingdu xuebao Vol. 28; no. 3; p. 231
Main Authors Xu, Peng-Wei, Guo, Jian-Qiang, Yao, Li-Hong, Chen, Ai-Jun, Liu, Xiao-Yu, Zeng, Xian-Yin, Zhang, Zhi-Qing
Format Journal Article
LanguageChinese
Published China 01.05.2012
Subjects
Animals
Baculoviridae - genetics
Baculoviridae - metabolism
Cell Line
Cloning, Molecular
Gene Expression
Genetic Vectors - genetics
Genetic Vectors - metabolism
Hemagglutinin Glycoproteins, Influenza Virus - genetics
Hemagglutinin Glycoproteins, Influenza Virus - immunology
Influenza A Virus, H1N1 Subtype - genetics
Influenza A Virus, H1N1 Subtype - immunology
Spodoptera
Transfection
Viral Matrix Proteins - genetics
Viral Matrix Proteins - immunology
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Summary: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.
ISSN:1000-8721