Stephens E B, Aulakh G S, Rose D L, Tully J G, Barile M F
Yale J Biol Med. 1983 Sep-Dec;56(5-6):729-35.
Radiolabeled DNA probes prepared in vitro by the nick translation method were used to determine the nucleotide sequence homology among the eight established and one unclassified species of Acholeplasma. Very little DNA homology (2 to 21 percent) was found among these nine distinct species and the heteroduplexes showed at least 15 percent mismatching as determined by thermal elution endpoints. The data obtained by hybridization analyses paralleled the results obtained by the growth inhibition and epi-immunofluorescence serologic procedures. The small amount of nucleotide sequence homology among the nine distinct species indicate that the Acholeplasma species are quite distinct and unrelated to each other genomically, findings which should provide useful insight on the molecular biology and evolutionary pathways of these organisms. Labeled 3H-DNA probes to five strains of either A. laidlawii or A. axanthum hybridized to a varying degree to excess amounts of unlabeled DNAs from 12 strains of A. laidlawii and six strains of A. axanthum, respectively. Nucleic acid hybridization analyses showed a wide variation (48 to 100 percent) in DNA homologies among different strains of the two species. The results demonstrate that strains of A. laidlawii and/or A. axanthum isolated from diverse hosts and habitats (birds, rodents, cats, swine, sheep, cattle, horses, goats, primates, and plants) exhibit extensive genotypic variations. 3H-DNA-DNA hybridization procedures were found to be extremely useful in establishing or confirming the existence of distinct species within the genus Acholeplasma.
通过缺口平移法体外制备的放射性标记DNA探针,用于确定8种已确定的和1种未分类的无胆甾原体物种之间的核苷酸序列同源性。在这9个不同的物种中发现的DNA同源性非常低(2%至21%),并且通过热洗脱终点测定,异源双链体显示至少15%的错配。杂交分析获得的数据与生长抑制和间接免疫荧光血清学方法获得的结果相似。这9个不同物种之间少量的核苷酸序列同源性表明,无胆甾原体物种在基因组上彼此非常不同且不相关,这些发现应为这些生物体的分子生物学和进化途径提供有用的见解。标记的3H-DNA探针与5株莱氏无胆甾原体或黄色无胆甾原体菌株分别与来自12株莱氏无胆甾原体和6株黄色无胆甾原体的过量未标记DNA进行了不同程度的杂交。核酸杂交分析表明,这两个物种的不同菌株之间的DNA同源性存在很大差异(48%至100%)。结果表明,从不同宿主和栖息地(鸟类、啮齿动物、猫、猪、绵羊、牛、马、山羊、灵长类动物和植物)分离的莱氏无胆甾原体和/或黄色无胆甾原体菌株表现出广泛的基因型变异。发现3H-DNA-DNA杂交程序在确定或确认无胆甾原体属内不同物种的存在方面极其有用。