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Expression and biological activity assay of bovine interleukin-18 fusion protein

Published online by Cambridge University Press:  27 June 2008

Tian Zhao-Ju ,
Zheng Yu-Shu ,
Liu Cui-Yan ,
Hu Jing-Dong  and
Zhao Hong-Kun
Tian Zhao-Ju
Affiliation:
The Faculty of Laboratory Medicine, Taishan Medical University, Tai'an 271016, China College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Tai'an 271018, China
Zheng Yu-Shu
Affiliation:
College of Animal Science, Henan Science College, Xinxiang 453003, China
Liu Cui-Yan
Affiliation:
College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Tai'an 271018, China
Hu Jing-Dong
Affiliation:
College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Tai'an 271018, China
Zhao Hong-Kun*
Affiliation:
College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Tai'an 271018, China
*
*Corresponding author. E-mail: [email protected]
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Abstract

The cDNA of bovine interleukin-18 (BoIL-18) was subcloned into pGEX6P-1 vector and transformed into Escherichia coli BL21(DE3). The recombinant protein was successfully expressed in E. coli by induction of isopropyl β-d-1-thiogalactopyranoside (IPTG) at 0.3 mmol/l for 8 h. SDS-PAGE indicated that the BoIL-18 fusion protein, 44 kDa, was highly expressed. Densitometric scanning showed that the fusion protein expression was about 31.8% of the total bacterial protein. The biological activity of the chromatographically purified protein was assayed. The peripheral blood mononuclear cells (PBMC) proliferation test indicated that the BoIL-18 fusion protein could enhance PBMC proliferation when its concentration was more than 0.10 mg/l. Enzyme-linked immunosorbent assay (ELISA) showed that the BoIL-18 fusion protein could induce interferon (IFN)-γ production from spleen lymphocytes when it was at a concentration of more than 0.20 mg/l, and that the inducing effect of BoIL-18 fusion protein on IFN-γ was directly proportional to its concentration. This verified that the purified BoIL-18 fusion protein possessed a functional activity and could be applied in further studies.

Keywords

biological activity assaybovine interleukin-18prokaryotic expressionpurification
Type
Research Papers
Information
Chinese Journal of Agricultural Biotechnology , Volume 5 , Issue 1 , April 2008 , pp. 7 - 11
Copyright
Copyright © China Agricultural University 2008

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Footnotes

First published in Journal of Agricultural Biotechnology 2007, 15(4): 579–583

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