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International Journal of Current Microbiology and Applied Sciences (IJCMAS)
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National Academy of Agricultural Sciences (NAAS)
NAAS Score: *5.38 (2019)
[Effective from January 1, 2019]
For more details click here

ICV 2017: 100.00
Index Copernicus ICI Journals Master List 2017 - IJCMAS--ICV 2017: 100.00
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 2017: 100.00
NAAS RATING 2018: 5.38

Int.J.Curr.Microbiol.App.Sci.2019.8(1): 1984-1990
DOI: https://doi.org/10.20546/ijcmas.2019.801.208


Expression and Purification of Recombinant Immunogenic Proteins of Goat Poxvirus in Prokaryotic System
Amit Kumar*1, Gnanavel Venkatesan1, Anand Kushwaha1, P. Sasi Kumar1, M.A. Ramakrishnan1 and Pronab Dhar2
1Division of Virology, ICAR- Indian Veterinary Research Institute, Mukteswar, Uttarakhand, India
2Division of Biological Standardization, ICAR-Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India
*Corresponding author
Abstract:

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 pET-33b(+) 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 capripoxvirus infections.


Keywords: Capripoxvirus, Expression, F13L protein, Goatpox virus, P32 protein, Prokaryotic, Western blot
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How to cite this article:

Amit Kumar, Gnanavel Venkatesan, Anand Kushwaha, P. Sasi Kumar, M.A. Ramakrishnan and Pronab Dhar. 2019. Expression and Purification of Recombinant Immunogenic Proteins of Goat Poxvirus in Prokaryotic System.Int.J.Curr.Microbiol.App.Sci. 8(1): 1984-1990. doi: https://doi.org/10.20546/ijcmas.2019.801.208