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Production of polyclonal viperin antisera using N-terminal deleted recombinant bovine viperin

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Title Production of polyclonal viperin antisera using N-terminal deleted recombinant bovine viperin
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Creator Sravanthi M, Sebastian R, Krishnaswamy N, Mahadappa P, Dechamma HJ, Umapathi V, Sanyal A
 
Subject Bovine viperin; cloning; expression; N-deletion; hyper immune serum
 
Description Not Available
Viperin, also known as radical S-adenosyl methionine domain-containing protein (RSAD2) is
a multifunctional interferon-stimulated gene (ISG) that is activated during the viral infections.
Viperin belongs to S-adenosyl methionine (SAM) superfamily of enzymes known to
catalyze radical-mediated reactions and viperin inhibits a wide range of DNA and RNA
viruses through its broad range of activity. The present study reports cloning and expression
of bovine viperin in a bacterial expression system. PCR-based site-directed mutagenesis was
carried out for deletion of N-terminal 1–70 amino acid containing amphipathic helix of
viperin that interferes in protein expression and purification. The resultant truncated viperin
protein was expressed in Escherichia coli, BL-21(DE3) competent cells and purified using
nickel charged affinity column. The truncated 54 kDa protein was confirmed by western blot
using human RSAD2 as a probe. Further, in house, hyperimmune serum was raised against
the truncated viperin in the rabbit and the reactivity was confirmed by western blot using
mammalian expression vector construct of viperin transfected in Baby Hamster kidney (BHK)
cells and in MDBK cells infected with Foot and Mouth disease Asia I virus
ICAR-IVRI
 
Date 2023-05-02T05:49:38Z
2023-05-02T05:49:38Z
2022-09-01
 
Type Research Paper
 
Identifier Mannem Sravanthi, Renjith Sebastian, Narayanan Krishnaswamy, Priyanka Mahadappa, H. J. Dechamma, V. Umapathi & Aniket Sanyal (2022): Production of polyclonal viperin antisera using N-terminal deleted recombinant bovine viperin, Animal Biotechnology, DOI: 10.1080/10495398.2022.2120890
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http://krishi.icar.gov.in/jspui/handle/123456789/76919
 
Language English
 
Relation Not Available;
 
Publisher TAYLOR FRANCIS