TY - JOUR
T1 - Isolation of oncogenes from rat mammary tumors by a highly efficient retrovirus expression cloning system
AU - Tsukamoto, Tetsuya
AU - Huang, Tony
AU - Guzman, Raphael C.
AU - Chen, Xiaoyan
AU - Pascual, Rhett V.
AU - Kitamura, Toshio
AU - Nandi, Satyabrata
N1 - Funding Information:
This work was supported by NIH Grants CA-05388 and CA-63369.
PY - 1999/11/11
Y1 - 1999/11/11
N2 - A majority of mammary tumors induced with N-methyl-N-nitrosourea in rats contain G to A transitional mutation of c-Ha-ras at the 12th codon. Additional oncogene activation is known to be necessary for further tumor progression. To isolate novel oncogenes, we used an expression cloning system utilizing the pMX retroviral vector in combination with BOSC23 packaging cells. First, we elucidated the sensitivity of this system in the NIH 3T3 focus assay; foci were detectable even after 10-6 dilution using v-Ha-ras, neuT, and β-galactosidase constructs in pMX vector. This system is sensitive enough to detect low copy number cDNAs. We used the pMX/BOSC23 expression cloning system to clone novel oncogenes from rat mammary tumors harboring an activated c-Ha ras and isolated several candidate oncogenes that caused transformation of NIH 3T3 cells and/or generated tumors when transplanted to nude mice.
AB - A majority of mammary tumors induced with N-methyl-N-nitrosourea in rats contain G to A transitional mutation of c-Ha-ras at the 12th codon. Additional oncogene activation is known to be necessary for further tumor progression. To isolate novel oncogenes, we used an expression cloning system utilizing the pMX retroviral vector in combination with BOSC23 packaging cells. First, we elucidated the sensitivity of this system in the NIH 3T3 focus assay; foci were detectable even after 10-6 dilution using v-Ha-ras, neuT, and β-galactosidase constructs in pMX vector. This system is sensitive enough to detect low copy number cDNAs. We used the pMX/BOSC23 expression cloning system to clone novel oncogenes from rat mammary tumors harboring an activated c-Ha ras and isolated several candidate oncogenes that caused transformation of NIH 3T3 cells and/or generated tumors when transplanted to nude mice.
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U2 - 10.1006/bbrc.1999.1625
DO - 10.1006/bbrc.1999.1625
M3 - Article
C2 - 10548482
AN - SCOPUS:0033547470
SN - 0006-291X
VL - 265
SP - 7
EP - 12
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -