Ortiz T, Piñero J, Cortés F
Department of Cell Biology, Faculty of Biology, University of Seville, Spain.
Mutat Res. 1995 Mar;327(1-2):161-9. doi: 10.1016/0027-5107(94)00183-6.
The possible recombination between non-homologous termini produced by restriction enzymes (REs) introduced in CHO cells by electroporation was studied. For this purpose, different combinations of REs that produced blunt or 5' overhanging DNA double-strand breaks were electroporated into cells either at the same time or separately by double electroporation experiments. Prior to double electroporation, it was confirmed that, once the cells have been electroporated, they resist a second electroporation, as assessed by cell viability analysis. Besides, the efficient and homogeneous introduction of labelled, non-permeable molecules was assessed by fluorescence microscopy. Our results showed interaction for most of the conditions, mainly when the REs were introduced separately. Differences found in the degree of interaction between the combinations studied are discussed.
研究了通过电穿孔导入中国仓鼠卵巢(CHO)细胞的限制性内切酶(REs)产生的非同源末端之间可能的重组。为此,通过双电穿孔实验,将产生平端或5'突出端DNA双链断裂的不同RE组合同时或分别电穿孔导入细胞。在双电穿孔之前,通过细胞活力分析证实,一旦细胞被电穿孔,它们就能抵抗第二次电穿孔。此外,通过荧光显微镜评估了标记的、不可渗透分子的有效和均匀导入。我们的结果表明,在大多数情况下都存在相互作用,主要是当REs分别导入时。讨论了在所研究的组合之间发现的相互作用程度的差异。