title: Expression of bovine (Bos indicus) interleukin-18 in Escherichia coli and its biological activity.
abstract: IL-18 modulates immune functions by inducing IFN-γ production and promoting Th1 immune responses. In the present study, we amplified and cloned the sequence (582 bp) encoding full-length bovine IL-18 from PBMC stimulated with PHA. The nucleotide and the deduced amino acid sequence of Bos indicus IL-18 showed an identity of 86-98% compared with IL-18 sequences of other ruminants. The insert was subcloned into a pET 32a vector and expressed in Escherichia coli as a fusion protein and the matured protein was obtained by caspase I treatment. The specificity of these proteins was confirmed by western blotting. The biological activity of the purified protein was analyzed by its ability to induce IFN-γ production in PBMC measured by ELISA and qPCR.
authors.affiliation: Indian Veterinary Research Institute, Bangalore-560024, Karnataka, India.
authors.forename: Kotla Siva
authors.initials: KS
authors.lastname: Reddy
authors.forename: Dowlathabad Muralidhar
authors.initials: DM
authors.lastname: Rao
authors.forename: Hosur Joyappa
authors.initials: HJ
authors.lastname: Dechamma
authors.forename: Veluvarthy V S
authors.initials: VV
authors.lastname: Suryanarayana
authors.forename: Golla Ramalinga
authors.initials: GR
authors.lastname: Reddy
mesh_terms: D000818:Animals; D002417:Cattle; D004926:Escherichia coli / Q000235:genetics*; D007371:Interferon-gamma / Q000096:biosynthesis; D020382:Interleukin-18 / Q000235:genetics* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000494:pharmacology
chemical_list: D020382:Interleukin-18; D011994:Recombinant Proteins; D007371:Interferon-gamma