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PMID: 2989794 Published · ppublish English Journal Article

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

Nucleic acids research ·Vol. 13 ·No. 12 ·1985-06-25 ·Pages 4417-29

Fransen L, Müller R, Marmenout A, Tavernier J, Van der Heyden J, Kawashima E, Chollet A, Tizard R, Van Heuverswyn H, Van Vliet A

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.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Cell Line Cloning, Molecular DNA/metabolism DNA Restriction Enzymes Dactinomycin/pharmacology Glycoproteins/genetics,physiology Growth Inhibitors/genetics L Cells/cytology Mice Plasmids Transcription, Genetic Tumor Necrosis Factor-alpha
Chemicals
Glycoproteins Growth Inhibitors Tumor Necrosis Factor-alpha Dactinomycin DNA DNA Restriction Enzymes
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Fransen L
Müller R
Marmenout A
Tavernier J
Van der Heyden J
Kawashima E
Chollet A
Tizard R
Van Heuverswyn H
Van Vliet A
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/13.12.4417
Published
1985-06-25
Pages
4417-29
Language
English
Region
England
NLM ID
0411011
PMCID
PMC321797
Subset
IM
Databases
GENBANK
X02611
External Links
PubMed source DOI: 10.1093/nar/13.12.4417 PMC Full Text View journal details
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