Ranhand J M
J Bacteriol. 1980 May;142(2):568-80. doi: 10.1128/jb.142.2.568-580.1980.
Competent cell-deoxyribonucleic acid (DNA) interactions were examined using tritium-labeled homologous or heterologous native or denatured DNAs and competent Streptococcus sanguis Wicky cells (strain WE4). The DNAs used were extracted from WE4 cells, Escherichia coli B cells, and E. coli bacteriophages T2, T4, T6, and T7. The reactions examined were: (i) total DNA binding, (ii) deoxyribonuclease-resistant DNA binding, and (iii) the production of acid-soluble products from the DNA. Optimal temperatures for the reactions were as follows: reaction (i), between 30 and 40 degrees C; reaction (ii), 30 degrees C; and reaction (iii), greater than 40 degrees C. The rates for the reactions (expressed as molecules of DNA that reacted per minute per colony-forming unit) did not vary greatly from one DNA source to another. With a constant competent cell concentration and differing DNA concentrations below a saturation level (from a given source), a different but constant fraction of the added DNA was cell bound, deoxyribonuclease resistant, and degraded to acid-soluble products. In experiments where the number of competent cells was varied and the DNA concentration was held constant, again essentially the same result was obtained. The extent of reactions (i), (ii), and (iii) depended upon the numbers as well as the source of DNA molecules applied to competent cells. Calcium ion essential for native DNA-cell reactions was also found essential for denatured DNA-cell reactions. Data obtained from competition experiments lead to the conclusion that competent WE4 cells contain specific sites for native as well as denatured DNAs.
使用氚标记的同源或异源天然或变性脱氧核糖核酸(DNA)以及感受态血链球菌Wicky细胞(菌株WE4)检测了感受态细胞与DNA的相互作用。所用的DNA提取自WE4细胞、大肠杆菌B细胞以及大肠杆菌噬菌体T2、T4、T6和T7。检测的反应包括:(i)总DNA结合,(ii)抗脱氧核糖核酸酶的DNA结合,以及(iii)DNA产生酸溶性产物。这些反应的最佳温度如下:反应(i),30至40摄氏度之间;反应(ii),30摄氏度;反应(iii),高于40摄氏度。各反应速率(以每分钟每个集落形成单位反应的DNA分子数表示)在不同DNA来源之间变化不大。在感受态细胞浓度恒定且DNA浓度低于饱和水平(来自给定来源)的情况下,添加的DNA中有不同但恒定比例的部分与细胞结合、抗脱氧核糖核酸酶且降解为酸溶性产物。在感受态细胞数量变化而DNA浓度保持恒定的实验中,再次得到了基本相同的结果。反应(i)、(ii)和(iii)的程度取决于应用于感受态细胞的DNA分子数量及其来源。发现天然DNA - 细胞反应所必需的钙离子对于变性DNA - 细胞反应同样必需。从竞争实验获得的数据得出结论,感受态WE4细胞含有天然DNA和变性DNA的特异性位点。