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| Expression and Analysis of H5N1 Avian Influenza Virus Hemagglutinin in Baculovirus Expression System |
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Abstract Abstract: HA gene of H5N1 avian influenza virus was amplified from the plasmid pGEM–HA and inserted into baculovirus transfer plasmid pFastBacHT to construct recombinant transfer vector pFastBacHT-HA. The pFastBacHT-HA was transformed into E.coli DH10Bac competent cells , transposited with baculovirus shuttle vector (Bacmid) and constructed recombinant transposition rBacmid-HA. After the rBacmid-HA transfected into sf9 cells , the recombinant baculovirus was harvested. The expressed HA protein was analyzed by SDS-PAGE, Western-blot and hemadsorption assay. The specific protein band of 66kDa was identified, which can only react with chicken sera against H5 subtype AIV and not react with the sera against H7 and H9 subtype respectively. Hemadsorption assay showed that sf9 cells that infected rBacmid-HT baculoviws can absorb chicken red blood cells. These results indicated that the HA protein was accurately expressed in sf9 cells, displayed nice specificity to H5 subtype antisera and biologic activation.
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Received: 19 December 2006
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