Phenotypic conversion of TK-deficient cells following electroporation of functional TK enzyme.
Phenotypic conversion of TK-deficient cells following electroporation of functional TK enzyme.
复制标题
功能性 TK 酶电穿孔后 TK 缺陷细胞的表型转化。
DOI：
10.1016/0014-4827(92)90146-y
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发表时间：
1992
影响因子：
3.7
通讯作者：
Patterson,RJ
中科院分区：
文献类型：
--
作者：
Dagher,SF;Conrad,SE;Werner,EA;Patterson,RJ
研究了通过电穿孔功能性TK酶来挽救突变(大鼠-3，胸苷激酶缺陷)细胞系的能力。电穿孔细胞的提取物显示，在90%的细胞仍然存活的条件下，TK酶水平增加了35倍。电穿孔的酶是细胞内的，事实证明，细胞能够利用外源[~3H]胸腺嘧啶核苷来合成DNA。在放射自显影中，82%的电穿孔细胞含有功能酶并将[~3H]胸腺嘧啶核苷掺入DNA。因此，这项技术可以有效地为培养的哺乳动物细胞提供缺失的代谢功能。
The ability to phenotypically rescue a mutant (Rat-3, thymidine kinase-deficient) cell line by electroporation of functional TK enzyme has been investigated. Extracts of electroporated cells showed a 35-fold increase in TK enzyme levels under conditions where >90% of the cells remained viable. The electroporated enzyme was intracellular, as demonstrated by the fact that cells were able to utilize exogenous [3H]thymidine for DNA synthesis. Byin situautoradiography, 82% of electroporated cells contained functional enzyme and incorporated [3H]thymidine into DNA. Thus, this technique can efficiently provide a missing metabolic function to cultured mammalian Cells.