Miller C C, McPheat J C, Potts W J
Biotechnology Department, ICI Pharmaceuticals, Macclesfield, Cheshire, United Kingdom.
Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):5020-4. doi: 10.1073/pnas.89.11.5020.
We have introduced a Ren-1D targeting vector into embryonic stem cells containing the two highly homologous mouse renin genes Ren-1D and Ren-2. Using a polymerase chain reaction (PCR) screen designed to detect targeted integration at Ren-1D and Ren-2, we isolated 15 targeted embryonic stem cell clones, all of which had undergone a gene conversion event at the Ren-1D locus. We did not isolate any clones in which the incoming DNA had recombined with Ren-2. Over the region encompassed by our transgene, Ren-1D and Ren-2 display greater than 95% homology. Our results suggest that the machinery driving gene targeting by means of homologous recombination in mammalian cells is capable of distinguishing between these two sequences. Construction of transgenic mice with the embryonic stem cells reported here carrying a mutated renin gene will permit a greater understanding of the functions of the Ren-1D and Ren-2 gene products and their relative contribution to cardiovascular homeostasis.
我们已将一种肾素-1D靶向载体导入含有两个高度同源的小鼠肾素基因肾素-1D和肾素-2的胚胎干细胞中。通过设计用于检测在肾素-1D和肾素-2处的靶向整合的聚合酶链反应（PCR）筛选，我们分离出15个靶向胚胎干细胞克隆，所有这些克隆在肾素-1D基因座都经历了基因转换事件。我们没有分离出任何导入的DNA与肾素-2发生重组的克隆。在我们的转基因所涵盖的区域，肾素-1D和肾素-2显示出大于95%的同源性。我们的结果表明，驱动哺乳动物细胞中通过同源重组进行基因靶向的机制能够区分这两个序列。用这里报道的携带突变肾素基因的胚胎干细胞构建转基因小鼠，将有助于更深入地了解肾素-1D和肾素-2基因产物的功能及其对心血管稳态的相对贡献。
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