Detection of transforming genes by transfection of DNA from primary soft-tissue tumours.
A series of adult soft-tissue tumours were screened for the presence of activated oncogenes by transfecting tumour DNA into NIH3T3 mouse fibroblasts. In these studies an activated K-ras gene that contained a mutation at the second position of codon 12 (GGT----GAT) was found in a leiomyosarcoma. In addition, following transfection of DNA from a liposarcoma, we identified an activated gene that failed to hybridize to probes prepared from 10 known human oncogenes (K-ras, H-ras, N-ras, ret, met, trk, mas, dbl, raf and hst) that have previously been detected in DNA transfection experiments. BamHI-BamHI fragments of this activated gene of 3.5 and 8.5 kb were cloned from NIH3T3 secondary transfectants using a probe that detects the human alu family of highly repetitive DNA sequences. Repeat-free subclones of these BamHI fragments were used to map this gene to human chromosome 19 (p13.2-q13.3). Our studies also demonstrate that a subclone from one of the BamHI fragments detects a 3.0 kb transcript in primary and secondary transfectants.
Section of Molecular Carcinogenesis, Institute of Cancer Research, Sutton, Surrey, UK.
S
S
Gill
M R
MR
Stratton
H
H
Patterson
N K
NK
Spurr
C
C
Fisher
B A
BA
Gusterson
C S
CS
Cooper
D016475:3T3 Cells; D000328:Adult; D000818:Animals; D002471:Cell Transformation, Neoplastic; D002888:Chromosomes, Human, Pair 19; D003001:Cloning, Molecular; D004273:DNA, Neoplasm / Q000235:genetics*; D006801:Humans; D006822:Hybrid Cells / Q000166:cytology; D008080:Liposarcoma / Q000235:genetics*; D051379:Mice; D009857:Oncogenes; D015183:Restriction Mapping; D012983:Soft Tissue Neoplasms / Q000235:genetics*; D014158:Transcription, Genetic; D014162:Transfection
D004273:DNA, Neoplasm