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 |
The σB protein of avian reovirus (ARV) is diagnostically important protein for development of an ARV-specific enzyme-linked immunosorbent assay (ELISA) and for which production of an antigenic recombinant σB protein in abundance is a prerequisite. In the present study, we successfully employed the prokaryotic system for expression of antigenic σB protein of ARV. The ARV isolated from the cases of viral arthritis and propagated on chicken embryo fibroblast primary culture was used for isolation of viral RNA. Isolated viral RNA was employed as a template for amplification of σB encoding gene. Further, the amplified fragment was cloned and subcloned in TA and pET32a(+) vectors, respectively. The continuity of open reading frame (ORF) was confirmed by sequenceing. The developed pET32a(+)-σB expression construct was transformed and expressed in BL21 (DE3) Rosetta cells. The SDS-PAGE and Western blot results confirmed the abundant expression of 56 kDa σB protein at 4 hr post induction. The expressed protein was further confirmed to be avian reovirus specific by its immunoreactivity with ARV hyperimmune serum. Overall, the results of the current study showed that the prokaryotic system can be exploited for expression of recombinant σB protein from avian reoviruses.
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