Scambler P J, Law H Y, Williamson R, Cooper C S
Nucleic Acids Res. 1986 Sep 25;14(18):7159-74. doi: 10.1093/nar/14.18.7159.
The DNA probes met and pJ3.11 are derived from loci on chromosome seven that are closely linked to, and probably flanking, the gene mutation causing cystic fibrosis (CF). We have shown that mitotic chromosomes from the cell line MNNG-HOS, which contains an activated met oncogene, can induce morphological transformation of mouse NIH-3T3 cells. Southern analysis of isolated transfectant cell lines with cloned dispersed repetitive human DNA sequences as probes demonstrated that several lines of transformed NIH 3T3 cells had stabley incorporated large segments of chromosome seven DNA. Southern blot analysis also demonstrated the presence of met, pJ3.11 and several other single copy sequences that had been previously localised to chromosome 7 within the transgenomes. In this way a further four genetic markers were shown to be physically linked to met, and thus to CF. These probes may prove useful in confirming the order of loci around CF and in the prenatal diagnosis of this common autosomal recessive disease.
DNA 探针 met 和 pJ3.11 源自七号染色体上的位点,这些位点与导致囊性纤维化(CF)的基因突变紧密连锁,可能位于该基因突变的两侧。我们已经证明,来自含有激活的 met 癌基因的细胞系 MNNG-HOS 的有丝分裂染色体,能够诱导小鼠 NIH-3T3 细胞发生形态转化。用克隆的分散重复人类 DNA 序列作为探针,对分离的转染细胞系进行 Southern 分析表明,几株转化的 NIH 3T3 细胞系已稳定整合了大片段的七号染色体 DNA。Southern 印迹分析还证明,在转基因组中存在 met、pJ3.11 以及其他几个先前已定位到七号染色体的单拷贝序列。通过这种方式,又有四个遗传标记被证明与 met 物理连锁,从而与 CF 连锁。这些探针可能有助于确定 CF 周围位点的顺序,并用于这种常见常染色体隐性疾病的产前诊断。