title: Identification by a random linker insertion method of a region which suppresses the transforming activity of the raf oncogene.
abstract: Activation mechanism of raf oncogene was studied by applying in vitro mutagenesis to its cDNA. Previous studies suggested the presence of an activation suppressing sequence in the amino-terminal half of c-raf product. Loss of the sequence by genetic rearrangement was presumed to convert c-raf to possess transforming activity. To identify such sequence, we prepared cDNA mutants by random linker insertion. Synthetic oligonucleotide linker was inserted into the plasmid containing cDNA at a single and random site. Coupling two different mutants, in-frame deletion mutants were constructed systematically. Analysis of these deletion mutants revealed a region, the loss of which made c-raf activated.
authors.affiliation: Carcinogenesis Division, National Cancer Center Research Institute, Tokyo, Japan.
authors.forename: F
authors.initials: F
authors.lastname: Ishikawa
authors.forename: R
authors.initials: R
authors.lastname: Sakai
authors.forename: M
authors.initials: M
authors.lastname: Ochiai
authors.forename: T
authors.initials: T
authors.lastname: Sugimura
authors.forename: M
authors.initials: M
authors.lastname: Nagao
mesh_terms: D000818:Animals; D002471:Cell Transformation, Neoplastic; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D004926:Escherichia coli / Q000235:genetics; D009154:Mutation; D009857:Oncogenes; D011519:Proto-Oncogenes; D051381:Rats; D013489:Suppression, Genetic
chemical_list: D004251:DNA Transposable Elements