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中国仓鼠卵巢细胞中的DNA介导基因转移:转移效率的克隆变异

DNA-mediated gene transfer in Chinese hamster ovary cells: clonal variation in transfer efficiency.

作者信息

Nairn R S, Adair G M, Humphrey R M

出版信息

Mol Gen Genet. 1982;187(3):384-90. doi: 10.1007/BF00332616.

Abstract

Thymidine kinase-deficient Chinese hamster ovary (CHO) cells were genetically transformed with the BamHI restriction fragment encoding the thymidine kinase gene of herpes simplex virus (HSV-tk). We have observed considerable clonal variation among independent CHO sublines with respect to transformation competence for the DNA-mediated gene transfer of HSV-tk. Transformation frequencies greater than or equal to 3 X 10(-4) were observed consistently in one subline, with a transformation efficiency of approximately 1 transformant per ng viral gene. The frequency and efficiency of transformation we observed in this system are at least 10-fold greater than those previously reported for DNA-mediated transformation of CHO cells by HSV-tk. All of the CHO HSV-tk+ transformants examined were stable for the transferred genotype in the absence of selection, and all showed evidence of co-transformation by unselected plasmid pBR322 sequences.

摘要

将编码单纯疱疹病毒胸苷激酶基因(HSV - tk)的BamHI限制性片段对胸苷激酶缺陷型中国仓鼠卵巢(CHO)细胞进行基因转化。我们观察到,在独立的CHO亚系中,对于HSV - tk的DNA介导基因转移,转化能力存在相当大的克隆变异。在一个亚系中始终观察到转化频率大于或等于3×10⁻⁴,转化效率约为每纳克病毒基因产生1个转化体。我们在该系统中观察到的转化频率和效率比先前报道的HSV - tk对CHO细胞进行DNA介导转化的频率和效率至少高10倍。在没有选择压力的情况下,所有检测的CHO HSV - tk⁺转化体对于转移的基因型都是稳定的,并且所有转化体都显示出未选择的质粒pBR322序列共转化的证据。

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