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| Cloning and Expression of Autographa californica nucleopolyhedrovirus orf59, Antibody Preparation and Its Subcellular Localization |
| Ling-Ling LI |
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Abstract Autographa californica nucleopolyhedrovirus (AcMNPV) orf59 gene was amplified from AcMNPV genomic DNA via PCR. The PCR product was cloned into the expression vector pET-32a(+) and transformed into Escherichia coli BL21, and a fusion protein around 30kDa was expressed after induction with IPTG (isopropylthio-β-D-galactoside).Purified fusion protein was injected into New Zealand white rabbit to harvest polyclonal antibodies. Western blotting analysis of extracts of AcMNPV-infected Sf9 cells showed two specific polypeptides with apparent sizes of 38kDa and 25kDa for the Ac59 antiserum. The indirect immunofluoresecence analysis demonstrated that the Ac59 protein was distributed in cytoplasm and nucleus of infected cells at the late stage of infection. This study lays a foundation for the further research of the gene function.
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Received: 03 September 2007
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Corresponding Authors:
Ling-Ling LI
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