Expression of bovine (Bos indicus) interleukin-18 in Escherichia coli and its biological activity.
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.
Reddy
Kotla Siva
KS
Indian Veterinary Research Institute, Bangalore-560024, Karnataka, India.
Rao
Dowlathabad Muralidhar
DM
Dechamma
Hosur Joyappa
HJ
Suryanarayana
Veluvarthy V S
VV
Reddy
Golla Ramalinga
GR
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
D020382:Interleukin-18; D011994:Recombinant Proteins; D007371:Interferon-gamma