通过表达原核二氢叶酸还原酶的重组质粒将小鼠成纤维细胞转化为甲氨蝶呤抗性。
Transformation of mouse fibroblasts to methotrexate resistance by a recombinant plasmid expressing a prokaryotic dihydrofolate reductase.
作者信息
O'Hare K, Benoist C, Breathnach R
出版信息
Proc Natl Acad Sci U S A. 1981 Mar;78(3):1527-31. doi: 10.1073/pnas.78.3.1527.
A recombinant plasmid has been constructed for the expression of inserted DNA sequences coding for polypeptide chains using the simian virus 40 early promoter and splicing and polyadenylylation signals from the rabbit beta-globin gene. The coding regions for two prokaryotic methotrexate-resistant dihydrofolate reductases were introduced into the expression vector. When mouse fibroblasts were exposed to these recombinant plasmids, it was possible to select methotrexate-resistant clones that had integrated the plasmids and produced a chimeric RNA coding for the prokaryotic enzyme.
已构建了一种重组质粒，用于利用猿猴病毒40早期启动子以及兔β-珠蛋白基因的剪接和聚腺苷酸化信号来表达编码多肽链的插入DNA序列。将两种原核抗甲氨蝶呤二氢叶酸还原酶的编码区导入表达载体。当小鼠成纤维细胞暴露于这些重组质粒时，有可能筛选出整合了质粒并产生编码原核酶的嵌合RNA的抗甲氨蝶呤克隆。
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