Expression of bovine (Bos indicus) interleukin-18 in Escherichia coli and its biological activity

Kotla Siva Reddy; Muralidhar Rao; Hosur Joyappa Dechamma; Veluvarthy V.S. Suryanarayana; Golla Ramalinga Reddy

doi:10.1111/j.1348-0421.2010.00241.x

Expression of bovine (Bos indicus) interleukin-18 in Escherichia coli and its biological activity

Kotla Siva Reddy, Muralidhar Rao, Hosur Joyappa Dechamma, Veluvarthy V.S. Suryanarayana, Golla Ramalinga Reddy

Research output: Contribution to journal › Article › peer-review

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.

Original language	English (US)
Pages (from-to)	564-567
Number of pages	4
Journal	Microbiology and Immunology
Volume	54
Issue number	9
DOIs	https://doi.org/10.1111/j.1348-0421.2010.00241.x
State	Published - Sep 2010
Externally published	Yes

Keywords

Biological activity
Bos indicus
Expression
Interleukin (IL-18)

ASJC Scopus subject areas

Microbiology
Immunology
Virology

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10.1111/j.1348-0421.2010.00241.x

Cite this

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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.",

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T1 - Expression of bovine (Bos indicus) interleukin-18 in Escherichia coli and its biological activity

AU - Reddy, Kotla Siva

AU - Rao, Muralidhar

AU - Dechamma, Hosur Joyappa

AU - Suryanarayana, Veluvarthy V.S.

AU - Reddy, Golla Ramalinga

PY - 2010/9

Y1 - 2010/9

N2 - 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.

AB - 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.

KW - Biological activity

KW - Bos indicus

KW - Expression

KW - Interleukin (IL-18)

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Expression of bovine (Bos indicus) interleukin-18 in Escherichia coli and its biological activity

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

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