Isolation of oncogenes from rat mammary tumors by a highly efficient retrovirus expression cloning system.
Isolation of oncogenes from rat mammary tumors by a highly efficient retrovirus expression cloning system.
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通过高效逆转录病毒表达克隆系统从大鼠乳腺肿瘤中分离癌基因。
DOI：
10.1006/bbrc.1999.1625
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
Nandi,S
中科院分区：
文献类型：
--
作者：
Tsukamoto,T;Huang,T;Guzman,RC;Chen,X;Pascual,RV;Kitamura,T;Nandi,S
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 10−6dilution 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.
