甲氨蝶呤抗性二氢叶酸还原酶基因在小鼠转化造血细胞中的表达。
Expression of a methotrexate-resistant dihydrofolate reductase gene by transformed hematopoietic cells of mice.
作者信息
Bar-Eli M, Stang H D, Mercola K E, Cline M J
出版信息
Somatic Cell Genet. 1983 Jan;9(1):55-67. doi: 10.1007/BF01544048.
DNA-mediated gene transfer was used to introduce DNA from a methotrexate-resistant mouse fibroblast cell line into mouse bone marrow cells. This cell line contained a methotrexate-resistant dihydrofolate reductase, active at 10(-4) M methotrexate, which was electrophoretically separable from the wild-type mouse enzyme. Transformed hematopoietic cells were returned to irradiated mice and selected in vivo by methotrexate administration. Some recipients of transformed marrow cells expressed the electrophoretically distinct, methotrexate-resistant dihydrofolate reductase in hematopoietic cells. These observations suggest that successful transformation of marrow stem cells to methotrexate resistance is accomplished by insertion of a dihydrofolate reductase gene coding for a mutant enzyme that is highly resistant to methotrexate.
利用DNA介导的基因转移技术，将来自耐甲氨蝶呤的小鼠成纤维细胞系的DNA导入小鼠骨髓细胞。该细胞系含有一种耐甲氨蝶呤的二氢叶酸还原酶，在10^(-4) M甲氨蝶呤浓度下仍具有活性，且在电泳中可与野生型小鼠酶分离。将转化的造血细胞回输到经照射的小鼠体内，并通过给予甲氨蝶呤在体内进行筛选。一些接受转化骨髓细胞的受体在造血细胞中表达出电泳特性不同的耐甲氨蝶呤二氢叶酸还原酶。这些观察结果表明，骨髓干细胞成功转化为耐甲氨蝶呤是通过插入一个编码对甲氨蝶呤高度耐药的突变酶的二氢叶酸还原酶基因来实现的。
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