Stanton V P, Cooper G M
Mol Cell Biol. 1987 Mar;7(3):1171-9. doi: 10.1128/mcb.7.3.1171-1179.1987.
Three activated cellular raf genes have been detected by transfection of NIH 3T3 cells with human tumor DNAs. Blot hybridization analysis indicated that all three transforming raf genes had recombined with non-raf sequences in the vicinity of raf exon 7-intron 7, resulting in the deletion of about 40% of the normal coding sequence from the raf amino terminus. By cloning sequences upstream of the truncated raf loci we have shown that the rearrangements involve the fusion of three different 5' non-raf human sequences to the human raf gene. No rearrangements could be detected in the raf loci of the three original human tumor DNAs, suggesting that the raf genes were activated by DNA rearrangements occurring during transfection. Significant overexpression of raf mRNA was not evident in two of the three transformant lines, indicating that raf overexpression is not necessary and 5' truncation alone may be sufficient to activate the transforming potential of cellular raf genes.
通过用人肿瘤DNA转染NIH 3T3细胞，已检测到三个激活的细胞raf基因。印迹杂交分析表明，所有三个转化raf基因均已与raf外显子7 - 内含子7附近的非raf序列重组，导致raf氨基末端约40%的正常编码序列缺失。通过克隆截短的raf基因座上游的序列，我们已表明重排涉及三种不同的5'非raf人类序列与人类raf基因的融合。在三个原始人类肿瘤DNA的raf基因座中未检测到重排，这表明raf基因是由转染过程中发生的DNA重排激活的。在三个转化细胞系中的两个中，raf mRNA的显著过表达并不明显，这表明raf过表达不是必需的，仅5'截短可能足以激活细胞raf基因的转化潜能。
