Chan T S, Long C, Green H
Somatic Cell Genet. 1975 Jan;1(1):81-90. doi: 10.1007/BF01538733.
A new selective medium has been developed for cells containing the enzyme deoxycytidine deaminase. This medium contains hypoxanthine, aminopterin, and 5-methyldeoxycytidine (HAM medium). To survive in the presence of the aminopterin, the cells must utilize deoxycytidine deaminase to convert the 5-methyldeoxycytidine to thymidine. The cells must also have thymidine kinase and hypoxanthine phosphoribosyltransferase. A mouse cell line deficient in deoxycytidine deaminase has been isolated from a deoxycytidine kinase-deficient line, using 5-bromodeoxycytidine as the selective agent. A hybrid line between this double mutant and a human diploid fibroblast was isolated in HAM medium. The hybrid line contains the chromosomes expected of a human-mouse hybrid. The deoxycytidine deaminase isozyme patterns on cellogel show that the human-mouse hybrid cell line produces an enzyme with an electrophoretic mobility intermediate between that of the human and that of the mouse.
已开发出一种针对含有脱氧胞苷脱氨酶的细胞的新型选择性培养基。这种培养基含有次黄嘌呤、氨基蝶呤和5-甲基脱氧胞苷(HAM培养基)。为了在氨基蝶呤存在的情况下存活,细胞必须利用脱氧胞苷脱氨酶将5-甲基脱氧胞苷转化为胸苷。细胞还必须具有胸苷激酶和次黄嘌呤磷酸核糖转移酶。利用5-溴脱氧胞苷作为选择剂,从缺乏脱氧胞苷激酶的细胞系中分离出了一种缺乏脱氧胞苷脱氨酶的小鼠细胞系。在HAM培养基中分离出了这种双突变体与人类二倍体成纤维细胞之间的杂交细胞系。该杂交细胞系含有预期的人鼠杂交染色体。细胞凝胶上的脱氧胞苷脱氨酶同工酶模式表明,人鼠杂交细胞系产生的一种酶的电泳迁移率介于人和小鼠的酶之间。