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Expression and purification of recombinant immunogenic proteins of goat poxvirus in prokaryotic system
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Capripox viruses of small ruminants, namely goatpox virus (GTPV) and sheep pox virus (SPPV) are responsible for important contagious diseases that are enzootic to the Indian sub-continent, Africa and the Middle East. In the present study, recombinant F13L and P32 proteins of GTPV were expressed in prokaryotic system, purified and confirmed in Western blot in order to evaluate their diagnostic potential. Full length F13L (1M-L370aa) and truncated P32 (20V-S270aa) genes of GTPV-Uttarkashi strain were cloned into pET33b(+) vector, over-expressed in prokaryotic system and purified as histidine-tagged protein using Ni-NTA affinity chromatography under denaturing conditions and passive elution method, respectively. The recombinantF13Land P32 proteins lacked fusion tag from vector except histidine tag for purification as analyzed by SDS-PAGE. Expression was confirmed with Western blot using anti-GTPV serum. The purified recombinant F13L and P32 proteins can be used potential diagnostic antigen/s either individually or in combination for sero-diagnosis of capripox virus infections. | Expression and purification of recombinant immunogenic proteins of goat poxvirus in prokaryotic system