National Academy of Agricultural Sciences (NAAS)
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PRINT ISSN : 2319-7692
Online ISSN : 2319-7706 Issues : 12 per year Publisher : Excellent Publishers Email : editorijcmas@gmail.com / submit@ijcmas.com Editor-in-chief: Dr.M.Prakash Index Copernicus ICV 2018: 95.39 NAAS RATING 2020: 5.38 |
Goatpox and sheeppox are highly contagious, OIE notifiable and economically important viral infections of small ruminants caused by goatpox virus (GTPV) and sheep poxvirus (SPPV) of genus Capripoxvirus, family Poxviridae. The disease is mainly endemic in central and northern Africa, Middle East and Asia including the Indian subcontinent. Several proteins of GTPV have been demonstrated to be immunogenic in nature. Out of these, G7L is a highly conserved major core protein encoded by ORF057 of GTPV genome and immunogenic in nature. The present study was designed to express full length G7L protein of GTPV using baculovirus expression vector system (BEVS) in insect cells and evaluate its diagnostic potential for detection of antibodies to GTPV. Initially, G7L gene of GTPV-Uttarkashi strain was amplified and cloned into BEVS donor vector, pFastBacTM HT A. The recombinant plasmid was confirmed by colony PCR/restriction enzyme analysis and transferred into DH10Bac cells containing baculovirus shuttle vector bacmid. The recombinant bacmid was isolated and transfected into Sf21 cells by using cellfectinTM reagent. Protein expression was analyzed by SDS-PAGE and confirmed by western blot as ~46 kDa in size. Recombinant G7L protein was purified by using Ni-NTA affinity chromatography under denaturing conditions and immunoreactivity of G7L protein was confirmed by using hyper immune serum of GTPV in western blot and indirect ELISA. The study confirmed immunoreactivity of recombinant G7L protein and further use as diagnostic reagent.
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