Transformation of NIH3T3 cells with synthetic c-Ha-ras genes.
Synthetic human c-Ha-ras genes in which amino acid codons were altered to those which are frequently used in highly expressed Escherichia coli genes were ligated to the 3'-end of Rous sarcoma virus long terminal repeat. When NIH3T3 cells were transfected with the plasmids having those genes with valine at codon 12, leucine at codon 61 or arginine at codon 61, transformants were efficiently produced. These results indicated that the synthetic c-Ha-ras genes are expressed in a mammalian system even though their codon usage is altered to correspond with that of E. coli. This expression vector system should be useful for studies on the structure-function relationships of c-Ha-ras, since the synthetic gene can be easily modified to have multiple base alterations, and can also be used simultaneously for the production of large amounts of p21 in E. coli for biochemical and biophysical studies.
Kamiya
H
H
Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo.
Miura
K
K
Ohtomo
N
N
Koda
T
T
Kakinuma
M
M
Nishimura
S
S
Ohtsuka
E
E
D001358:Avian Sarcoma Viruses / Q000235:genetics; D001483:Base Sequence; D002461:Cell Line, Transformed; D002471:Cell Transformation, Neoplastic; D003001:Cloning, Molecular; D003062:Codon; D004279:DNA, Viral / Q000235:genetics; D006801:Humans; D009154:Mutation; D010957:Plasmids; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D016283:Proto-Oncogene Proteins p21(ras); D011519:Proto-Oncogenes; D012091:Repetitive Sequences, Nucleic Acid; D014162:Transfection
D003062:Codon; D004279:DNA, Viral; D011518:Proto-Oncogene Proteins; C501009:HRAS protein, human; D016283:Proto-Oncogene Proteins p21(ras)