Huttner K M, Scangos G A, Ruddle F H
Proc Natl Acad Sci U S A. 1979 Nov;76(11):5820-4. doi: 10.1073/pnas.76.11.5820.
We have used DNA-mediated gene transfer to introduce a recombinant plasmid containing the human beta-globin gene (H beta 1) into cells of a mouse tissue culture line, Ltk-. DNA isolated from independent transfer lines was analyzed by restriction endonuclease digestion, gel electrophoresis, modified Southern blotting, and filter hybridization using H beta 1 as a probe. H beta 1 sequences were present in 80% of the lines at 1-30 copies per cell. Many of the lines gave a hybridization pattern indicative of H beta 1 sequences integrated into high molecular weight DNA. DNA from three cell lines, digested with several restriction enzymes, produced a pattern providing evidence for the presence of circular H beta 1 molecules in the murine recipient cells.
我们已利用DNA介导的基因转移，将一个含有人类β-珠蛋白基因（Hβ1）的重组质粒导入小鼠组织培养细胞系Ltk-中。通过限制性内切酶消化、凝胶电泳、改良的Southern印迹法以及使用Hβ1作为探针的滤膜杂交，对从独立转移系中分离出的DNA进行了分析。Hβ1序列以每个细胞1 - 30个拷贝的形式存在于80%的细胞系中。许多细胞系呈现出一种杂交模式，表明Hβ1序列已整合到高分子量DNA中。用几种限制性酶消化三个细胞系的DNA后，产生的模式为小鼠受体细胞中存在环状Hβ1分子提供了证据。
