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通过DNA介导的转化将基因转移到中国仓鼠卵巢细胞中。

Transfer of genes to Chinese hamster ovary cells by DNA-mediated transformation.

作者信息

Abraham I, Tyagi J S, Gottesman M M

出版信息

Somatic Cell Genet. 1982 Jan;8(1):23-39. doi: 10.1007/BF01538648.

Abstract

We have transferred DNa to Chinese hamster ovary (CHO) cells by DNA-mediated transformation. CHO tk- cells were transformed with the clones gene for herpes simplex virus thymidine kinase (HSV-tk) and were found to have a 50-fold lower frequency of transformation than mouse Ltk- cells at the same DNA dosage. By altering the amount of tk gene and carrier DNA present, frequencies of up to 5 x 10(-5) were obtained. CHO HSV-tk+ transformants were very stable, and in several clones the HSV-tk gene copies integrated in higher-molecular-weight DNA. These cells also exhibited cotransformation for unselected markers. CHO lines were also transformed at a frequency of 10(-4) with the bacterial gene Ecogpt in a SV40-pBR322 vector. CHO tk-cells could be transformed at a frequency of 10(-7) with cellular DNA isolated from CHO tk+ cells. CHO cells offer a well-defined genetic system within which to transfer either cloned or whole cellular DNAs.

摘要

我们通过DNA介导的转化将DNA导入了中国仓鼠卵巢(CHO)细胞。用单纯疱疹病毒胸苷激酶(HSV-tk)的克隆基因对CHO tk-细胞进行转化,发现在相同DNA剂量下,其转化频率比小鼠Ltk-细胞低50倍。通过改变tk基因和载体DNA的量,获得了高达5×10⁻⁵的频率。CHO HSV-tk⁺转化体非常稳定,在几个克隆中,HSV-tk基因拷贝整合到了高分子量DNA中。这些细胞还表现出对未选择标记的共转化。用SV40-pBR322载体中的细菌基因Ecogpt也能以10⁻⁴的频率转化CHO细胞系。用从CHO tk⁺细胞中分离的细胞DNA能以10⁻⁷的频率转化CHO tk-细胞。CHO细胞提供了一个定义明确的遗传系统,可在其中导入克隆的或完整的细胞DNA。

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