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用可扩增的显性作用基因转化哺乳动物细胞。

Transformation of mammalian cells with an amplifiable dominant-acting gene.

作者信息

Wigler M, Perucho M, Kurtz D, Dana S, Pellicer A, Axel R, Silverstein S

出版信息

Proc Natl Acad Sci U S A. 1980 Jun;77(6):3567-70. doi: 10.1073/pnas.77.6.3567.

Abstract

We have transferred a mutant hamster gene coding for an altered dihydrofolate reductase to wild-type cultured mouse cells by using total genomic DNA from methotrexate-resistant Chinese hamster ovary A29 cells as donor. By demonstrating the presence of hamster gene sequences in transformants we have provided direct evidence for gene transfer. Transformants selected for increased resistance to methotrexate contain increased amounts of the newly transferred gene. We have used this mutant dhfr gene to introduce the Escherichia coli antibiotic resistance plasmid pBR322 into animal cells. Amplification of the dhfr sequences results in amplification of the pBR322 sequences as well. The use of this gene may allow the introduction and amplification of virtually any genetic element in various new cellular environments.

摘要

我们通过使用来自甲氨蝶呤抗性中国仓鼠卵巢A29细胞的全基因组DNA作为供体,将编码改变的二氢叶酸还原酶的突变仓鼠基因转移到野生型培养的小鼠细胞中。通过证明转化体中存在仓鼠基因序列,我们为基因转移提供了直接证据。选择对甲氨蝶呤抗性增加的转化体含有增加量的新转移基因。我们已经使用这个突变的二氢叶酸还原酶基因将大肠杆菌抗生素抗性质粒pBR322引入动物细胞。二氢叶酸还原酶序列的扩增也导致pBR322序列的扩增。使用这个基因可能允许在各种新的细胞环境中引入和扩增几乎任何遗传元件。

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