EXPRESSION OF ABBREVIATED MOUSE DIHYDROFOLATE-REDUCTASE GENES IN CULTURED HAMSTER-CELLS
EXPRESSION OF ABBREVIATED MOUSE DIHYDROFOLATE-REDUCTASE GENES IN CULTURED HAMSTER-CELLS
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DOI：
10.1073/pnas.79.21.6522
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发表时间：
1982-01-01
期刊：
影响因子：
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通讯作者：
SCHIMKE, RT
中科院分区：
文献类型：
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作者：
GASSER, CS;SIMONSEN, CC;SCHIMKE, RT
利用克隆的小鼠基因组和互补DNA序列构建了一些二氢叶酸还原酶（DHFR）基因的缩写形式。这些基因包含1.0千碱基的5′侧翼基因组序列以及3′非编码区的不同部分。其中两个基因包含DHFR基因的前两个内含子；另外三个则缺乏内含子。用这些构建的任何一种基因转染二氢叶酸还原酶缺陷型的中国仓鼠卵巢细胞，都会产生具有二氢叶酸还原酶阳性表型的细胞。用甲氨蝶呤（一种叶酸拮抗剂）处理转染细胞，会导致出现对甲氨蝶呤有抗性的集落，这些集落会扩增所转染的基因。
A number of abbreviated dihydrofolate reductase (DHFR) genes were constructed by using cloned mouse genomic and c[complementary]DNA sequences. These genes contain 1.0 kilobase of 5'' flanking genomic sequence and varying portions of the 3'' non-coding region. Two of the genes contain the first 2 introns of the DHFR gene; the other 3 lack introns. Transfection of DHFR-deficient Chinese hamster ovary cells with any of these constructed genes results in cells with the DHFR+ phenotype. Treatment of the transfectants with methotrexate, a folate antagonist, leads to the emergence of methotrexate-resistant colonies which amplified the transfected genes.