Hostname: page-component-cd9895bd7-7cvxr Total loading time: 0 Render date: 2024-12-27T09:57:30.252Z Has data issue: false hasContentIssue false
  • English
  • Français

Expression of H5N1 Avian influenza virus haemagglutinin in a baculovirus expression system

Published online by Cambridge University Press:  13 February 2008

Dun Jifeng ,
Pu Juan ,
Zhou Yingchun  and
Liu Jinhua
Dun Jifeng
Affiliation:
Department of Infectious Disease, College of Veterinary Medicine, China Agricultural University, Beijing 100094, China
Pu Juan
Affiliation:
Department of Infectious Disease, College of Veterinary Medicine, China Agricultural University, Beijing 100094, China
Zhou Yingchun
Affiliation:
Department of Infectious Disease, College of Veterinary Medicine, China Agricultural University, Beijing 100094, China
Liu Jinhua*
Affiliation:
Department of Infectious Disease, College of Veterinary Medicine, China Agricultural University, Beijing 100094, China
*
*Corresponding author. E-mail: ljh@cau.edu.cn
Get access Rights & Permissions [Opens in a new window]

Abstract

The haemagglutinin (HA) gene of H5N1 Avian influenza virus (AIV) was amplified from the plasmid pGEM-HA and cloned into the baculovirus transfer plasmid pFastBacHT to construct the recombinant transfer vector pFastBacHT-HA. The pFastBacHT-HA was transformed into DH10Bac competent cells, transposed with a shuttle vector (Bacmid) and the transposition rBacmid-HA was constructed. The recombinant baculovirus was harvested from sf9 cells transfected with rBacmid-HA. The expressed HA protein was identified and analysed by SDS–PAGE, Western blot and haemadsorption assay. The 66 kDa protein could only react with chicken serum positive to H5 subtype AIV. The haemadsorption assay showed that the sf9 cells infected with rBacmid-HA baculovirus could absorb chicken red blood cells. These results indicated that the HA protein was successfully expressed in sf9 cells, with reaction specificity to H5 subtype antiserum.

Keywords

Avian influenza virusH5N1haemagglutininbaculovirus expression system
Type
Research Article
Information
Chinese Journal of Agricultural Biotechnology , Volume 4 , Issue 3 , December 2007 , pp. 233 - 237
Copyright
Copyright © China Agricultural University and Cambridge University Press 2007

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

Footnotes

†

These authors contributed equally to this work.

First published in Journal of Agricultural Biotechnology 2007, 15(4): 547–551

References

Gan, MH (2004) Avian Influenza, 2nd ed. Beijing: China Agricultural Press (in Chinese).Google Scholar
Gao, YS and Wang, C (2006) Analysis of global avian influenza epidemic situation in 2006. Journal of Inspection and Quarantine 2: 3–8 (in Chinese).Google Scholar
Jackson, DC, Murray, JM, White, DO and Gerhard, WU (1982) Enumeration of antigenic site of influenza virus hemagglutinin. Infection and Immunity 37: 912–918.CrossRefGoogle ScholarPubMed
Lee, CW, Suarez, DL, Tumpey, TM, et al. (2005) Characterization of highly pathogenic H5N1 avian influenza A viruses isolated from south Korea. Journal of Virology 79: 3692–3702.CrossRefGoogle ScholarPubMed
Liu, JH, Xiao, HX, Lei, FM, et al. (2005) Highly pathogenic H5N1 influenza virus infection in migratory birds. Science 309: 1206.CrossRefGoogle ScholarPubMed
Nakajima, K, Nobusawa, E, Tonegawa, K and Nakajima, S (2003) Restriction of amino acid change in H3HA: Comparision of amino acid changes observed in nature and in vitro. Journal of virology 77(18): 10088–10098.CrossRefGoogle Scholar
Nakajima, K, Nobusawa, E, Nagy, A and Nakajima, S (2005) Accumulation of amino acid substitutions promotes irreversible structural changes in the hemagglutinin of human influenza A H3 virus during evolution. Journal of Virology 79: 6472–6477.CrossRefGoogle Scholar
Ohuchi, M, Fischer, C, Ohuchi, R, Herwig, A and Klenk, HD (1998) Elongation of the cytoplasmic tail interferes with the fusion activity of influenza virus hemagglutinin. Journal of Virology 72(5): 3554–3595.CrossRefGoogle ScholarPubMed
Stevens, J, Corper, AL, Basler, CF, Taubenberger, JK, Palese, P and Wilson, IA (2004) Structure of the uncleaved human H1 hemagglutinin from the extinct 1918 influenza virus. Science 303: 1866–1869.CrossRefGoogle ScholarPubMed
Stevens, J, Blixt, O, Tumpey, MT, Jeffery, KT, Paulson, JC and Wilson, IA (2006a) Structure and receptor specificity of the hemmagglutinin from an H5N1 influenza virus. Science 312: 404–410.CrossRefGoogle Scholar
Stevens, J, Blixt, O, Glaser, L, et al. (2006b) Glycan microarray analysis of the hemagglutinins from modern and pandemic influenza viruses reveals different receptor specificities. Journal of Molecular Biology 355: 1143–1155.CrossRefGoogle ScholarPubMed
Wilson, IA, Skehel, JJ and Wiley, DC (1981) Structure of the haemagglutinin membrane glycoprotein of influenza virus at 3Å resolution. Nature 289: 366–373.CrossRefGoogle Scholar
Wu, S, Chen, C, Lei, T, Cao, H and Chen, FY (2005) Expression of hemagglutinin gene of avian influenza virus subtype H5 in Escherichia coli and the preliminary analysis of bioactivity. Journal of Agricultural Biotechnology 13(6): 739–742 (in Chinese).Google Scholar
Zeng, Z and Wu, DY (2005) Progress of Baculovirus expression vector system. Journal of Sericulture in China 26(3): 4–7 (in Chinese).Google Scholar