Capsid protein genes vp2 and vp3 of Taura syndrome virus (TSV) were cloned into pet-16b-1 and pGEX-4t-3 expression vector respectively, and transformed intoEscherichia coli BL21 or DH5α for protein expression and purification. After induction with IPTG, recombinant VP2 (rVP2) and recombinant VP3 (rVP3) were produced in E. coli, purified by SDS-PAGE and used to immunize Balb/c mice forthe production of polyclonal antisera: anti-rVP2 and anti-rVP3. Two antigenic peptides originating from TSV capsid protein VP2 and VP3 respectively were synthesized as antigen for the production of monoclonal antibodies (Mabs). Mab specific to VP2, VP3, anti-rVP2 and anti-rVP3 antisera all showed specific immunoreactivities to corresponding recombinant viral proteins by Western blot assay. The Mabs as well as the polyclonal antisera could detect VP2 and VP3 in homogenates from gills of TSV-infected shrimp by immunodot-blot. This is the first step towards our target of preparing TSV capsid proteins as oral vaccine and developing simple immuno-diagnostic test kits for TSV detection in Penaeus vannamei shrimp.
Key words: Taura syndrome virus (TSV), Penaeus vannamei, Mabs, VP2 and VP3 protein, Western blot.
TSV, Taura syndrome virus PCR, polymerase chain reaction; RT-PCR, reverse transcription polymerase chain reaction; SDS-PAGE, SDS polyacrylamide gel electrophoresis; Mab, monoclonal antibodies.
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