Molecular cloning of mouse tumour necrosis factor cDNA and its eukaryotic expression | Zendy

Lucie Fransen | Zendy; Rita Müller | Zendy; Anne Marmenout | Zendy; Jan Tavernier | Zendy; José Van der Heyden | Zendy; Eric Kawashima | Zendy; André Chollet | Zendy; Richard Tizard | Zendy; H. Van Heuverswyn | Zendy; Adri Van Vliet | Zendy; Marie-Rose Ruysschaert | Zendy; Walter Fiers | Zendy

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Molecular cloning of mouse tumour necrosis factor cDNA and its eukaryotic expression

Author(s) -

Lucie Fransen,

Rita Müller,

Anne Marmenout,

Jan Tavernier,

José Van der Heyden,

Eric Kawashima,

André Chollet,

Richard Tizard,

H. Van Heuverswyn,

Adri Van Vliet,

Marie-Rose Ruysschaert,

Walter Fiers

Publication year - 1985

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/13.12.4417

Subject(s) - biology , complementary dna , microbiology and biotechnology , transfection , recombinant dna , tumor necrosis factor alpha , oligonucleotide , amino acid , plasmid , messenger rna , molecular cloning , expression vector , cloning (programming) , peptide sequence , gene , cdna library , coding region , biochemistry , immunology , computer science , programming language

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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