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International Journal of Current Microbiology and Applied Sciences (IJCMAS)
IJCMAS is now DOI (CrossRef) registered Research Journal. The DOIs are assigned to all published IJCMAS Articles.
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National Academy of Agricultural Sciences (NAAS)
NAAS Score: *5.38 (2020)
[Effective from January 1, 2020]
For more details click here

ICV 2018: 95.39
Index Copernicus ICI Journals Master List 2018 - IJCMAS--ICV 2018: 95.39
For more details click here

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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

Int.J.Curr.Microbiol.App.Sci.2017.6(9): 434-441
DOI: https://doi.org/10.20546/ijcmas.2017.609.052


High-Yield Expression and Purification of Recombinant σB Protein of Avian Reovirus (ARV) in Prokaryotic System
Sanjeevna K. Minhas1*, Nitin M. Kamble1, J.M. Kataria2, C. Madhan Mohan1 and Sohini Dey1
1Recombinant-DNA Laboratory, Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, UP - 243122, India
2Central Avian Research Institute, Izatnagar, UP - 243122, India
*Corresponding author
Abstract:

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.


Keywords: Avian reovirus, σB protein, Expression, Prokaryotic.
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How to cite this article:

Sanjeevna K. Minhas, Nitin M. Kamble, J.M. Kataria, C. Madhan Mohan and Sohini Dey. 2017. High-Yield Expression and Purification of Recombinant σB Protein of Avian Reovirus (ARV) in Prokaryotic SystemInt.J.Curr.Microbiol.App.Sci. 6(9): 434-441. doi: https://doi.org/10.20546/ijcmas.2017.609.052