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

Published online by Cambridge University Press:  13 February 2008

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

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.

Type
Research Article
Copyright
Copyright © China Agricultural University and Cambridge University Press 2007

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Footnotes

These authors contributed equally to this work.

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

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