Molecular cloning of mouse tumour necrosis factor cDNA and its eukaryotic expression (original) (raw)

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Biogent, Jozef Plateaustraat 22, B-9000 Ghent, Belgium

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Biogent, Jozef Plateaustraat 22, B-9000 Ghent, Belgium

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Biogent, Jozef Plateaustraat 22, B-9000 Ghent, Belgium

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Biogent, Jozef Plateaustraat 22, B-9000 Ghent, Belgium

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Biogent, Jozef Plateaustraat 22, B-9000 Ghent, Belgium

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Biogen S.A., 46, Route des Acacias, CH-1227 Carouge/Geneva, Switzerland

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Biogen S.A., 46, Route des Acacias, CH-1227 Carouge/Geneva, Switzerland

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Biogen Inc., 14 Cambridge Center, Cambridge, MA 02142, USA

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Biogent, Jozef Plateaustraat 22, B-9000 Ghent, Belgium

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Biogent, Jozef Plateaustraat 22, B-9000 Ghent, Belgium

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Lucie Fransen, Rita Mūller, Anne Marmenout, Jan Tavernier, José Van der Heyden, Eric Kawashima, André Chollet, Richard Tizard, Hugo Van Heuverswyn, Adri Van Vliet, Marie-Rose Ruysschaert, Walter Fiers, Molecular cloning of mouse tumour necrosis factor cDNA and its eukaryotic expression, Nucleic Acids Research, Volume 13, Issue 12, 25 June 1985, Pages 4417–4429, https://doi.org/10.1093/nar/13.12.4417
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Abstract

Tumour necrosis factor (TNF), released by induced macrophages, causes tumour necrosis in animals and kills preferentially transformed cells in vitro. mRNA induced in the established mouse monocytic PU 5.1.8 cell line by lipopolysaccharide, was converted into double-stranded cDNA and cloned in the pAT153 vector. Recombinant plasmids were screened by plus-minus hybridization and TNF-specific oligonucleotide probes constructed on the basis of partial amino acid sequences of rabbit TNF. A series of TNF specific clones were identified and confirmed by hybrid selection of mouse TNF-specific mRNA. The sequence codes for a 235 amino acids long polypeptide, of which 156 amino acids presumably correspond to the mature product. It can be concluded that mature mouse TNF is a glycosylated dimer. Biologically active TNF was secreted by both Cos-I and CHO-cells transfected with the chimaeric expression vector pSV2d2-mTNF containing the coding region of the mouse TNF cDNA gene

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