PMID: 8960095Nov 29, 1996Paper

Interleukin-1 receptor antagonist: characterisation of its gene expression in rabbit tissues and large-scale expression in eucaryotic cells using a baculovirus expression system

Journal of Immunological Methods
J ApostolopoulosP G Tipping

Abstract

The gene expression of rabbit interleukin-1 receptor antagonist (RbIL-lra) was examined in rabbit tissues. RNA was isolated from heart, lung, kidney, muscle, liver, spleen, brain, and peripheral blood monocytes (PBMs), and RbIL-lra mRNA was identified as a single species by Northern analysis using a RbIL-lra probe. RbIL-lra was abundantly expressed in lung, brain, heart, and liver, expressed at low levels in spleen, and undetectable in kidney and unstimulated PBMs. Expression of large scale recombinant production of RbIL-lra was achieved by subcloning the cDNA into a baculovirus expression vector. Recombination of this vector was completed with the BacPAK6 baculovirus genome. The recombinant virus, containing the RbIL-lra cDNA, was used to infect Spodoptera frugiperda (Sf21) insect cells in a spinner flask system and in monolayers in cell culture flasks. Recombinant rabbit IL-lra (rRbIL-lra) was secreted into the culture medium in this system at very high levels (35 mg/l). The protein was identified by reducing SDS-PAGE electrophoresis, was variably glycosylated and had a molecular weight between 19-25 kDa. It was then purified by size exclusion HPLC on a Du Pont Gf-250 column. The rRbIL-lra was demonstrated to be functionally ...Continue Reading

References

Sep 15, 1978·Cellular Immunology·S B MizelJ J Oppenheim
Nov 1, 1991·Immunology Today·C A Dinarello, R C Thompson
Jul 1, 1983·Analytical Biochemistry·A P Feinberg, B Vogelstein
Apr 15, 1983·Cellular Immunology·B KleinB Serrou
Aug 1, 1996·Arteriosclerosis, Thrombosis, and Vascular Biology·J ApostolopoulosP G Tipping

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Citations

Nov 26, 1999·Journal of Interferon & Cytokine Research : the Official Journal of the International Society for Interferon and Cytokine Research·T TakiiK Onozaki
May 23, 1998·Annual Review of Immunology·W P ArendC Gabay
Mar 14, 2000·Immunity·J ApostolopoulosM S Sandrin
Aug 6, 2005·The Journal of Immunology : Official Journal of the American Association of Immunologists·Michelle N WykesMichael F Good
Jun 13, 2012·The Journal of Immunology : Official Journal of the American Association of Immunologists·Mark S GibsonPete Kaiser

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