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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 Mannem Sravanthi
Sebastian R
Krishnaswamy N
Mahadappa P
Dechamma HJ
Umapathi V
Sanyal A
 
Subject bovine viperin
cloning
expression
Ndeletion
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.
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Date 2023-05-10T04:17:22Z
2023-05-10T04:17:22Z
2022-09-16
 
Type Research Paper
 
Identifier Sravanthi M, Sebastian R, Krishnaswamy N, Mahadappa P, Dechamma HJ, Umapathi V, Sanyal A. Production of polyclonal viperin antisera using N-terminal deleted recombinant bovine viperin. Anim Biotechnol. 2022 Sep 16:1-8. doi: 10.1080/10495398.2022.2120890. Epub ahead of print. PMID: 36112063.
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http://krishi.icar.gov.in/jspui/handle/123456789/76999
 
Language English
 
Relation Not Available;
 
Publisher TAYLOR FRANCIS