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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.
Index Copernicus ICI Journals Master List 2019 - IJCMAS--ICV 2019: 96.39 For more details click here
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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 2019: 96.39
Index Copernicus ICI Journals Master List 2019 - IJCMAS--ICV 2019: 96.39
For more details click here

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Original Research Articles

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.2020.9(7): 1717-1724
DOI: https://doi.org/10.20546/ijcmas.2020.907.198


Baculovirus Expression and Immunoreactivity of G7L Core Protein of Goatpox Virus
Anand Kushwaha, Amit Kumar, S. Chandra Sekar, Golmei Poulinlu, Durga Goswami, Dhanavelu Muthuchelvan, Muthannan Andavar Ramakrishnan and Gnanavel Venkatesan*
Division of Virology, ICAR- Indian Veterinary Research Institute, Mukteswar, Uttarakhand-263138, India
*Corresponding author
Abstract:

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.


Keywords: Gaotpox virus, G7L, Core protein, Baculovirus expression vector system, Insect cell line, Western blot, Indirect ELISA
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How to cite this article:

Anand Kushwaha, Amit Kumar, S. Chandra Sekar, Golmei Poulinlu, Durga Goswami, Dhanavelu Muthuchelvan, Muthannan Andavar Ramakrishnan and Gnanavel Venkatesan. 2020. Baculovirus Expression and Immunoreactivity of G7L Core Protein of Goatpox Virus.Int.J.Curr.Microbiol.App.Sci. 9(7): 1717-1724. doi: https://doi.org/10.20546/ijcmas.2020.907.198