Huttner K M, Barbosa J A, Scangos G A, Pratcheva D D, Ruddle F H
J Cell Biol. 1981 Oct;91(1):153-6. doi: 10.1083/jcb.91.1.153.
DNA-mediated gene transfer is a procedure which uses purified DNA to introduce new genetic elements into cells in culture. The standard DNA-mediated gene transfer procedure involves the use of whole cell DNA as carrier DNA for the transfer. We have modified the standard DNA-mediated gene transfer procedure to transfer the Herpes simplex virus type 1 thymidine kinase gene (TK) into TK- murine recipient cells in the absence of whole cell carrier DNA. The majority (8/10) of carrier-free transformant lines expressed the TK+ phenotype stably, in sharp contrast to our results with carrier-containing DNA-mediated gene transfer. There was a wide range in donor DNA content among independent transformants. Further analysis on one transformant line using DNA restriction digests and in situ hybridization provided evidence that, in the absence of whole cell carrier DNA, multiple donor DNA sequences became integrated at a single chromosomal site.
DNA介导的基因转移是一种利用纯化的DNA将新的遗传元件导入培养细胞的方法。标准的DNA介导的基因转移程序涉及使用全细胞DNA作为转移的载体DNA。我们对标准的DNA介导的基因转移程序进行了改进，以便在没有全细胞载体DNA的情况下将单纯疱疹病毒1型胸苷激酶基因（TK）转移到TK-小鼠受体细胞中。大多数（8/10）无载体转化体系稳定地表达了TK+表型，这与我们使用含载体的DNA介导的基因转移的结果形成了鲜明对比。独立转化体之间的供体DNA含量差异很大。对一个转化体系使用DNA限制性酶切和原位杂交进行的进一步分析提供了证据，即在没有全细胞载体DNA的情况下，多个供体DNA序列在单个染色体位点整合。
