van der Lugt N, Maandag E R, te Riele H, Laird P W, Berns A
Division of Molecular Genetics, The Netherlands Cancer Institute, Amsterdam.
Gene. 1991 Sep 15;105(2):263-7. doi: 10.1016/0378-1119(91)90161-4.
We have constructed a hypoxanthine phosphoribosyl transferase-selectable marker (hprt) under the control of the phosphoglycerate kinase (pgk) promoter. This construct permits cell growth in hypoxanthine/aminopterin/thymidine media and confers 6-thioguanine sensitivity upon mouse Hprt- embryonic stem cells, allowing either positive or negative selection in gene-targeting experiments. We have successfully targeted the gene encoding the T-cell receptor delta-chain using the pgk::hprt fusion for counterselection.
我们构建了一个在磷酸甘油酸激酶（pgk）启动子控制下的次黄嘌呤磷酸核糖基转移酶选择标记（hprt）。该构建体允许细胞在次黄嘌呤/氨基蝶呤/胸腺嘧啶核苷培养基中生长，并赋予小鼠Hprt-胚胎干细胞对6-硫鸟嘌呤的敏感性，从而在基因靶向实验中实现正向或负向选择。我们已经成功地使用pgk::hprt融合体进行反选择，对编码T细胞受体δ链的基因进行了靶向。
