Mulsant P, Gatignol A, Dalens M, Tiraby G
Laboratoire de Génétique Cellulaire, INRA, Castanet-Tolosan, France.
Somat Cell Mol Genet. 1988 May;14(3):243-52. doi: 10.1007/BF01534585.
The Tn5 and the Streptoalloteichus hindustanus (Sh) ble genes conferring resistance to bleomycin-phleomycin antibiotics have been cloned into a mammalian vector under the RSV-LTR promoter. The resulting plasmids, pUT506 and pUT507 respectively, were used to transfect CHO cells by either the calcium phosphate or the recently described polybrene-DMSO method. Phleomycin- or bleomycin-resistant clones arose with a higher frequency after transfection with pUT507, and pUT507 transfectants were more resistant to both antibiotics than pUT506 transfectants. Phleomycin resistance in pUT507 transfectants was stable and associated with integration of plasmid sequences in genomic DNA. The Sh ble gene, which confers a dominant phleomycin-resistance phenotype, should provide a useful transferable selectable marker in CHO cells as well as in other animal cell lines.
赋予对博来霉素-腐草霉素类抗生素抗性的Tn5和印度斯坦链霉菌（Sh）ble基因已在劳斯肉瘤病毒长末端重复序列（RSV-LTR）启动子控制下克隆到一个哺乳动物载体中。所得到的质粒分别为pUT506和pUT507，通过磷酸钙法或最近描述的聚凝胺-二甲基亚砜法用于转染中国仓鼠卵巢（CHO）细胞。用pUT507转染后，博来霉素或腐草霉素抗性克隆出现的频率更高，并且pUT507转染子比pUT506转染子对这两种抗生素都更具抗性。pUT507转染子中的腐草霉素抗性是稳定的，并且与质粒序列整合到基因组DNA中有关。赋予显性腐草霉素抗性表型的Sh ble基因应在CHO细胞以及其他动物细胞系中提供一个有用的可转移选择标记。
