PMID: 7538248Feb 1, 1995Paper

Expression, biological activity and kinetics of production of recombinant ovine TNF-alpha

Veterinary Immunology and Immunopathology
H F SeowP Wood

Abstract

Ovine tumour necrosis factor-alpha (OvTNF-alpha) was cloned by reverse transcription-polymerase reaction using RNA isolated from lipopolysaccharide (LPS)-stimulated alveolar macrophages and primers based on the human TNF-alpha cDNA sequence. An expression vector carrying the coding sequence of the mature form of ovine TNF was constructed. The recombinant Ov-TNF alpha (rOvTNF-alpha) was expressed as a glutathione-S-transferase (GST) fusion protein. It was cleaved with thrombin to yield rOvTNF free of the GST moiety. Growth at a lower temperature of 30 degrees C and use of Escherichia coli strains AM207, AM305, E392 and NM522 did not improve the recovery of rOvTNF-alpha from the soluble fraction to a significant extent. Purification of recombinant proteins was achieved rapidly and easily by affinity chromatography using glutathione-Sepharose. Yields of pure rOvTNF-alpha achieved in E. coli JM109 and AM207 were approximately 1 mg L-1. Both rOvTNF-alpha and recombinant human TNF-alpha (rhTNF-alpha) exerted cytotoxicity on L929 cells. However, rOvTNF-alpha but not rhTNF-alpha stimulated proliferation of ovine thymocytes. Maximum levels of TNF-alpha mRNA expression by LPS-stimulated ovine alveolar macrophages were detected at approxi...Continue Reading

References

Feb 15, 1992·Veterinary Immunology and Immunopathology·A D NashM R Brandon
Oct 25, 1991·Proceedings of the National Academy of Sciences of the United States of America·L A TartagliaD V Goeddel
Aug 1, 1991·Immunology and Cell Biology·A D NashA E Andrews
Apr 1, 1991·Proceedings of the National Academy of Sciences of the United States of America·M LewisD V Goeddel
Nov 25, 1990·Nucleic Acids Research·A J YoungJ Y Chan
Jan 1, 1989·Annual Review of Immunology·B Beutler, A Cerami
Jan 1, 1988·Journal of Basic Microbiology·K SreekrishnaP J Wood
Feb 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·D S Silberstein, J R David
Jan 1, 1986·Cold Spring Harbor Symposia on Quantitative Biology·D V GoeddelB J Sugarman
Oct 24, 1986·Science·K J TraceyJ D Albert
Sep 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·D PennicaD V Goeddel

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Citations

Sep 1, 1996·Veterinary Immunology and Immunopathology·K PerssonH F Seow
Jun 5, 2004·Developmental and Comparative Immunology·Carol G Chitko-McKownWilliam W Laegreid
Jan 1, 2003·Research in Veterinary Science·K Persson WallerK Ostensson
Dec 1, 1999·Developmental and Comparative Immunology·S RautenschleinJ M Sharma
Nov 1, 1996·Veterinary Immunology and Immunopathology·P R Wood, H F Seow
May 1, 1997·Veterinary Immunology and Immunopathology·C W WongD L Watson
Jul 1, 1997·Veterinary Immunology and Immunopathology·J S RothelP R Wood
Jan 22, 1998·Veterinary Immunology and Immunopathology·C M DozoisI P Oswald
Feb 26, 2000·Veterinary Immunology and Immunopathology·P McWatersJ P Scheerlinck
Feb 7, 2006·Journal of Immunological Methods·Sangeeta BudhiaBarbara A Blacklaws
Aug 1, 1996·Immunology and Cell Biology·C W WongD L Watson

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