Molecular cloning and nucleotide sequence analysis of a transforming gene from human gastrocarcinoma cell line.
Mouse and rat fibroblasts were transfected with total DNA from human gastrocarcinoma cell line BGC-823. It was shown by hybridization assay that the genome of one of the rat secondary foci contains transforming genes from the human gastrocarcinoma cell line, which are homologous to the protooncogene c-Ha-ras in the normal cells. The genomic library of the rat secondary foci was constructed, using lambda phage EMBL3 as the vector. The transforming gene Ha-ras of the human gastrocarcinoma cell was thus cloned by screening the library with the probes of human Alu repeat sequence and c-Ha-ras. The nucleotide sequences of the first and second exons were analysed by M13-dideoxy method. The result shows that the nucleotide sequence of the transforming gene is the same as that of the normal protooncogene except one nucleotide difference in the first exon.
Deng
G R
GR
Beijing Institute for Cancer Research.
Lü
Y Y
YY
Chen
S M
SM
Miao
J
J
Lu
G R
GR
Li
H
H
Cai
H
H
Xu
X L
XL
Zheng
E
E
Liu
P N
PN
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D004273:DNA, Neoplasm / Q000235:genetics*; D005347:Fibroblasts; D011905:Genes, ras; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D051381:Rats; D013274:Stomach Neoplasms / Q000235:genetics* / Q000473:pathology; D014162:Transfection
D004273:DNA, Neoplasm