Hemagglutination and hemadsorption activities associated with S1 glycoprotein of infectious bronchitis virus
Date Issued
2012
Date
2012
Author(s)
Lin, You-Hsin
Abstract
Abstract
A Taiwan local infectious bronchitis virus strain 2575 TW-I showed hemagglutination activity after neuraminidase treatment while its attenuated strain did not. In order to investigate this altered hemagglutination activity, we cloned the S1 gene of the attenuated strain. The nucleotide differences between original and attenuated strains were mutated in the S1 gene by site direct mutation. The S1 gene in the pFastBacHT A was transferred to the bacmids in DH10Bac. The Bac-to-Bac baculovirus expressing system was used to express both wild and attenuated strain S1 proteins. After transfection and passages, the P3 stock of recombinant baculovirus was generated and purified by sucrose gradient separation. The hemagglutination (HA) activity and cell-associtaed hemadsorption (Hd) activity of the expression proteins were examined. Both recombinant baculoviruses showed Hd but no HA activities. The Hd activities can be inhibited by anti-IBV antiserum. Through this method, desired S1 protein mutant could be expressed with the baculovirus expressing system and further studied for biological activity and immunological reactivity.
Subjects
Hemadsorption
Hemagglutination
Infectious bronchitis virus
Type
thesis
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