Production of polyclonal viperin antisera using Nterminal deleted recombinant bovine viperin.
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Title |
Production of polyclonal viperin antisera using Nterminal deleted recombinant bovine viperin.
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Creator |
Mannem Sravanthi, Renjith Sebastian, Narayanan Krishnaswamy, Priyanka Mahadappa, H. J. Dechamma, V. Umapathi and Aniket Sanyal
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Subject |
Bovine viperin; cloning; expression; N-deletion; hyper immune serum
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Description |
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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. Not Available |
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Date |
2023-05-02T04:53:44Z
2023-05-02T04:53:44Z 2022-09-16 |
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Type |
Research Paper
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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.
Not Available http://krishi.icar.gov.in/jspui/handle/123456789/76908 |
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Language |
English
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Relation |
Not Available;
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Publisher |
Not Available
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