Production of antibodies against DNA polymerase of Bombyx mori nuclear polyhedrosis virus
1996
Sudawan Chaeychomsri | Ikeda, Motoko | Kobayashi, Michihiro (Kasetsart Univ. Kamphaengsaen Campus, Nakhon Pathom (Thailand). Central Laboratory and Greenhouse Complex Center)
DNA polymerase gene (pol) of Bombyx mori nuclear polyhedrosis virus (BmNPV) was inserted downstream from the malE gene of Escherichia coli, which encodes maltose-binding protein (MBP), resulting in the expression of an MBP-Pol fusion protein. The MBP-Pol fusion product was purified by affinity chromatography. The Pol fusion protein was used as the immunogen for the production of polyclonal antibodies. The specificity and titer of these antibodies were examined against MBP-Pol fusion protein and DNA polymerase in the infected cells by dot-immunobinding assay and by Western blot analysis, respectively. using these antibodies, the accumulation of BmNPV Pol protein in the fat body, midgut, silkgland and remaining carcass of the silkworm larvae infected with BmNPV was examined. The Western blot analysis showed the positive band of 125K protein which was not observed in the uninfected tissues. The apparent molecular weight (MW) of this protein was roughly comparable to 115K of the BmNPV DNA polymerase predicted from the open reading frame (ORF) of pol.
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