Jones I M, Burkhart-Schultz K, Crippen T L
Biomedical Sciences Division, Lawrence Livermore National Laboratory, California 94550.
Somat Cell Mol Genet. 1987 Jul;13(4):325-33. doi: 10.1007/BF01534926.
As part of our mouse model of somatic mutation, we have begun to characterize spontaneously occurring hypoxanthine phosphoribosyltransferase (HPRT) -deficient mouse lymphocytes. Lymphocytes were cloned by in vitro exposure of spleen cells from male C57B1/6 mice to the mitogen concanavalin A, conditioned medium containing lymphocyte growth factors, and thioguanine (TG), in a limiting dilution assay. The 17 TG-resistant clones recovered were all highly deficient in HPRT activity and were found by analysis of surface antigens to be representative of the major subclasses of T lymphocytes. Southern analysis of lymphocyte genomic DNA detected alterations of the hprt gene in 12/17 of the HPRT-deficient lymphocyte clones. Of these 12, 2/17 were lacking the entire hprt locus, 7/17 lacked part of the locus, and 3/17 had other, unidentified alterations.
作为我们体细胞突变小鼠模型的一部分，我们已开始对自发出现的次黄嘌呤磷酸核糖转移酶（HPRT）缺陷型小鼠淋巴细胞进行表征。通过在有限稀释试验中，将雄性C57B1/6小鼠的脾细胞体外暴露于促有丝分裂剂刀豆球蛋白A、含有淋巴细胞生长因子的条件培养基和硫鸟嘌呤（TG），对淋巴细胞进行克隆。回收的17个对TG有抗性的克隆在HPRT活性方面均高度缺陷，并且通过表面抗原分析发现它们代表了T淋巴细胞的主要亚类。对淋巴细胞基因组DNA的Southern分析在17个HPRT缺陷型淋巴细胞克隆中的12个中检测到hprt基因的改变。在这12个中，2/17缺失整个hprt基因座，7/17缺失部分基因座，3/17有其他未鉴定的改变。
