Porter B W, Kolodner R, Warner R C
J Bacteriol. 1973 Oct;116(1):163-74. doi: 10.1128/jb.116.1.163-174.1973.
The six plasmids of Shigella dysenteriae Y6R were separated by sucrose gradients into five fractions containing deoxyribonucleic acid (DNA), having contour lengths (expressed in units equal to the fraction of the length of the replicative form of phiX174), respectively, of 0.29, 0.35, 0.74, 1.08, and a mixture of 5.7 and 7.2. DNA-DNA hybridization on nitrocellulose filters between each of the plasmids and between plasmid-free S. dysenteriae Y6R host DNA and plasmids was investigated. There was a high degree of homology between the 0.29- and 0.35-unit plasmids. No significant homology was found between any of the other pairs of plasmids. Homologous DNA to the extent of 2.4 copies of the 1.08-unit plasmid was found in the host genome. Homology between the other plasmids and the host genome is very slight, but appears to be significant. About 0.7 of the 1.08-unit plasmid is homologous to the ColE1 façtor of Escherichia coli JC411 (ColE1). This plasmid may be defective ColE1 factor with the immunity function intact, but with a defect in the gene leading to the production of active colicin. Electron microscope examination of heteroduplexes formed between the two smallest plasmids and between the 1.08-unit plasmid and the ColE1 factor yielded independent determinations of the extent of homology in agreement with the values determined by hybridization. In the latter case, two nonhomologous regions of substitution of DNA were detected.
痢疾志贺氏菌Y6R的6个质粒通过蔗糖梯度分离成5个含有脱氧核糖核酸(DNA)的级分,其轮廓长度(以等于phiX174复制形式长度的分数为单位表示)分别为0.29、0.35、0.74、1.08,以及5.7和7.2的混合物。研究了每个质粒之间以及无质粒的痢疾志贺氏菌Y6R宿主DNA与质粒之间在硝酸纤维素滤膜上的DNA-DNA杂交。0.29单位和0.35单位的质粒之间存在高度同源性。在其他任何一对质粒之间均未发现明显的同源性。在宿主基因组中发现了与1.08单位质粒2.4份拷贝程度的同源DNA。其他质粒与宿主基因组之间的同源性非常轻微,但似乎很显著。1.08单位质粒的约0.7与大肠杆菌JC411的ColE1因子(ColE1)同源。该质粒可能是免疫功能完整但导致活性大肠杆菌素产生的基因有缺陷的ColE1因子。对两个最小质粒之间以及1.08单位质粒与ColE1因子之间形成的异源双链体进行电子显微镜检查,得到的同源性程度独立测定结果与杂交测定值一致。在后一种情况下,检测到两个DNA替代的非同源区域。