Identification by a random linker insertion method of a region which suppresses the transforming activity of the raf oncogene
Ishikawa, F.; Sakai, R.; Ochiai, M.; Sugimura, T.; Nagao, M.
Nucleic Acids Symposium Series 19: 39-42
1988
ISSN/ISBN: 0261-3166 PMID: 2852353 Document Number: 317457
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.