Kamiyama T, Aoki N, Yuasa Y
Department of Hygiene and Oncology, Tokyo Medical and Dental University School of Medicine.
Jpn J Cancer Res. 1993 Mar;84(3):250-6. doi: 10.1111/j.1349-7006.1993.tb02864.x.
cDNA clones presumably containing a transforming gene were isolated from transformed NIH3T3 cells, which were obtained by the transfection of DNA from peripheral blood lymphocytes of a familial adenomatous polyposis patient. Sequence analysis of the cDNA clones showed that the gene contained an activated B-raf, the 5' half of which was replaced by the human small nuclear ribonucleoprotein E protein gene and an unknown gene. A partial genomic physical map of the transforming gene was made on the basis of the physical map of the cDNA clones, indicating that rearrangements had occurred during the transfection. The human B-raf gene was shown to be located on chromosome 7 by Southern blotting analysis of rodent-human somatic cell hybrid DNA using the B-raf cDNA as a probe.
从转化的NIH3T3细胞中分离出可能含有转化基因的cDNA克隆,这些细胞是通过转染来自一名家族性腺瘤性息肉病患者外周血淋巴细胞的DNA获得的。对cDNA克隆的序列分析表明,该基因包含一个激活的B-raf,其5'端的一半被人类小核核糖核蛋白E蛋白基因和一个未知基因所取代。基于cDNA克隆的物理图谱构建了转化基因的部分基因组物理图谱,表明在转染过程中发生了重排。通过使用B-raf cDNA作为探针,对啮齿动物-人类体细胞杂种DNA进行Southern印迹分析,显示人类B-raf基因位于7号染色体上。