Tomilin N V, Titomirov A V, Stepan'ian L I, Dyban P A
Tsitologiia. 1987 Dec;29(12):1385-91.
A plasmid containing the bacterial gene of methotrexate-resistant dihydrofolate reductase (dhfr), under the control of early SV40 promoter, was introduced into murine teratocarcinoma CBA9H6 cells. From the whole pool of teratocarcinoma cells, which survived after transient methotrexate selection in vivo, the individual cells were isolated to give rise to 15 clones of tumors. Six of the 15 clones displayed nucleotide sequences of the original vector containing pBR322 sequences and the early SV40 promoter region; however, the bacterial dhfr gene was absent from the transformant clones. Possible causes of the loss of introduced dhfr gene from teratocarcinoma cells under non-selective conditions are discussed.
将一个含有抗甲氨蝶呤二氢叶酸还原酶(dhfr)细菌基因的质粒,置于早期SV40启动子的控制下,导入小鼠畸胎瘤CBA9H6细胞。在体内经短暂甲氨蝶呤筛选后存活的整个畸胎瘤细胞群体中,分离出单个细胞以产生15个肿瘤克隆。15个克隆中的6个显示出含有pBR322序列和早期SV40启动子区域的原始载体的核苷酸序列;然而,转化克隆中不存在细菌dhfr基因。讨论了在非选择性条件下畸胎瘤细胞中导入的dhfr基因丢失的可能原因。