Vatteroni L, Piras A, Simi S, Mariani L, Moretti A, Citti L, Mariani T, Rainaldi G
Istituto di Mutagenesi e Differenziamento, CNR, Pisa, Italy.
Mutat Res. 1993 Jun;291(3):163-9. doi: 10.1016/0165-1161(93)90156-t.
Electroporation is a recent technique used to introduce exogenous DNA into eukaryotic cells. It is important to establish that the gene of interest is transferred into a functional, non-mutated recipient cell. V79/AP4 Chinese hamster cells were exposed to high-voltage pulsed electric fields and some biological and genetic effects were measured. The results showed that cytotoxicity was related in a dose-dependent manner to the number of applied pulses. Thioguanine-resistant colony-forming cells as well as chromosomal aberrations were also induced whereas ouabain resistants and sister-chromatid exchanges were not or slightly induced. Spontaneous and electroporation-induced clones that were phenotypically TGR/HATS were used to investigate the hprt locus. Molecular screening of the locus showed that the number of deleted exons was significantly higher in induced than in spontaneous TG-resistant clones, suggesting that the genetic damages induced by electroporation concern the loss of regions well over the size of the hprt locus.
电穿孔是一种最近用于将外源DNA导入真核细胞的技术。确定感兴趣的基因被转移到一个功能正常、未发生突变的受体细胞中很重要。将V79/AP4中国仓鼠细胞暴露于高压脉冲电场，并测量了一些生物学和遗传学效应。结果表明，细胞毒性与施加的脉冲数呈剂量依赖性关系。还诱导出了硫代鸟嘌呤抗性集落形成细胞以及染色体畸变，而哇巴因抗性和姐妹染色单体交换未被诱导或仅轻微诱导。对表型为TGR/HATS的自发和电穿孔诱导克隆进行了hprt基因座研究。对该基因座的分子筛选表明，诱导的TG抗性克隆中缺失外显子的数量明显高于自发克隆，这表明电穿孔诱导的遗传损伤涉及远远超过hprt基因座大小的区域的丢失。
