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| Baculovirus Expression of Truncated HA Gene of A/H1N1 Influence Virus |
| JIA Yuan1,2, SHEN Yang1, QIU Ya-feng1, SHI Zi-xue1, SHAO Dong-hua1, WANG Xiao-du1, DENG Xu-Fang1, WANG Hao-ting1, ZHAO Fu-guang2, MA Zhi-yong1 |
1. Shanghai Veterinary Research Institute, The Chinese Academy of Agricultural Sciences, Shanghai 200237, China;
2. Jilin Agricultural University, Changchun 130118, China |
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Abstract Object: In order to express recombinant HA protein using the Bac-to-Bac Baculovirus Expression System. The expression of the recombinant HA protein was confirmed by Western blot and IFA. Method: A/California/04/2009(H1N1)HA gene was amplified by PCR, and the PCR product was cloned into the pFastBacHT A vector. The recombinant plasmid pFastBacHT-HA was identified by restriction enzyme digestion and gene sequencing. The recombinant plasmid pFastBacHT-HA was subsequently transformed into E. coli DH10Bac component cells. The recombinant rbacmid-HA identified by Bluo-gal blue/white selection and PCR was purified and transfected into sf 9 cells. The P1 virus obtained from sf 9 cells was amplified and stocked. The sf 9 cells infected with the baculovirus was harvested and analyzed for detection of the expression of recombinant protein by Western blot and IFA. Conclusion: The expression vector engineered for expressing A/H1N1 influence virus HA gene was constructed, which was transfected into insect cells, The recombinant baculovirus with higher viral titer was obtained. The recombinant protein was expressed by recombinant baculovirus in the infected sf 9 cell and identified by Western blot and IFA, demonstrating that the truncated HA protein can be recognized by the antibody specific to HA of 2009 H1N1 influenza virus.
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Received: 22 March 2011
Published: 25 October 2011
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Cite this article:
JIA Yuan, SHEN Yang, QIU Ya-feng, SHI Zi-xue, SHAO Dong-hua, WANG Xiao-du, DENG Xu-Fang, WANG Hao-ting, ZHAO Fu-guang, MA Zhi-yong. Baculovirus Expression of Truncated HA Gene of A/H1N1 Influence Virus. China Biotechnology, 2011, 31(10): 18-22.
URL:
https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2011/V31/I10/18
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