Shapira G, Stachelek J L, Letsou A, Soodak L K, Liskay R M
Proc Natl Acad Sci U S A. 1983 Aug;80(15):4827-31. doi: 10.1073/pnas.80.15.4827.
Thymidine kinase-deficient mouse L cells have been transformed with plasmid DNAs carrying 8-base-pair Xho I linker insertion mutations in the coding region of the herpes simplex virus type 1 thymidine kinase gene. When the mutant plasmids are introduced individually into LTK- cells, transformation efficiencies are greatly reduced relative to the wild type. However, when two mutant plasmids are cotransferred into the same LTK- recipients, significantly higher frequencies of transformation are observed (30-300 times). Here we demonstrate the usefulness of linker insertions for the study of homologous recombination in detecting the existence of normal thymidine kinase gene sequences (i.e., sequences lacking the insertions after recombination are substantiated by DNA . DNA hybridization). In addition, the frequencies of recombination in the various "crosses" are consistent with the known positions of the mutations.
胸苷激酶缺陷型小鼠L细胞已被携带1型单纯疱疹病毒胸苷激酶基因编码区8碱基对Xho I接头插入突变的质粒DNA转化。当将突变体质粒单独导入LTK⁻细胞时，相对于野生型，转化效率大大降低。然而，当将两个突变体质粒共转入同一LTK⁻受体细胞时，观察到显著更高的转化频率（30 - 300倍）。在这里，我们证明了接头插入在检测正常胸苷激酶基因序列存在（即重组后缺乏插入的序列通过DNA·DNA杂交得到证实）的同源重组研究中的有用性。此外，各种“杂交”中的重组频率与已知的突变位置一致。
