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Vet Immunol Immunopathol. 2008 Dec 15;126(3-4):403-6. doi: 10.1016/j.vetimm.2008.08.004. Epub 2008 Aug 23.

Expression, purification and characterisation of recombinant Escherichia coli derived chicken interleukin-12.

Author information

1
CSIRO - Australian Animal Health Laboratories, Livestock Industries, Private Bag 24, Geelong, Victoria 3220, Australia.

Abstract

Zoonotic viruses, such as H5N1 Avian Influenza, pose major threats to both animals and humans, and with this in mind there is a need for the development of new anti-viral strategies. The cytokine interleukin-12 (IL-12) is known to play a pivotal regulatory role in the anti-viral response due to its role in the induction of the key anti-viral cytokine IFN-gamma. Therefore, strategies which provide a means for the production of therapeutic quantities of IL-12 may be of major benefit. Here we describe the development of biologically active Escherichia coli (E. coli) derived chicken IL-12 (ChIL-12). The single chain ChIL-12 gene was cloned into the pET32b expression vector, transformed into the BL-21 E. coli strain and expression induced with IPTG. Over expressed protein was solubilised with zwittergent detergent and isolated utilising Nickel ion affinity chromatography. Biological activity was determined as ChIL-12 stimulated proliferation of pre-treated T-cells in vitro. This study is the first example of a biologically active E. coli derived IL-12 from a non-mammalian vertebrate subsequently providing a means for testing the anti-viral therapeutic potential of ChIL-12 in an in vivo model.

PMID:
18952299
DOI:
10.1016/j.vetimm.2008.08.004
[Indexed for MEDLINE]

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