1PhD Scholar, Indian Veterinary Research Institute, Hebbal, Bangalore-560024, Karnataka, India
2Principal Scientist, Indian Veterinary Research Institute, Hebbal, Bangalore-560024, Karnataka, India
*Corresponding author email id: umapathi_pillai@yahoo.co.in
Online published on 9 March, 2022.
The host response to viral infections includes stimulation of pattern recognition receptors which induces the release of type I interferons. Type I interferons further stimulate hundreds of interferons stimulated genes (ISGs). ISG15, a ubiquitin like protein, is an interferon stimulated gene implicated in antiviral response in several viral infections. Characterization of bovine ISG15 is important for studying the role of this ISG in pathogenesis of viral infections in cattle. Cloning and expression of the ISG15 in a bacterial expression system is the first step in characterization of this protein. Therefore, the present study is aimed at PCR amplification of the coding sequences of ISG15 using mRNA isolated from Foot and Mouth Disease Virus infected MDBK cells and to clone the amplified PCR product in pET32a to express in bacterial expression systems. The analysis of nucleotide and amino acid sequence of the cloned ISG15 revealed that it is highly conserved within bovine species. The cloned pET32a-ISG15 recombinant protein was successfully expressed in E. coli expression system and characterized by SDS-PAGE analysis.
ISG15, pET32a, Cloning, Expression
