Crawford J T, Cave M D, Bates J H
Rev Infect Dis. 1981 Sep-Oct;3(5):949-52. doi: 10.1093/clinids/3.5.949.
Strains of Mycobacterium avium-intracellulare were examined for the presence of plasmids. Actively growing cultures were treated with cycloserine and ampicillin to sensitize the cells to lysis by detergent, and cleared lysates were prepared by treatment with sodium dodecyl sulfate and NaCl. Purified plasmid DNA was prepared by equilibrium density centrifugation in CsCl gradients containing ethidium bromide and analyzed by agarose gel electrophoresis. Strain LR25 was found to carry three plasmids, including a large plasmid with a molecular weight of greater than 10(8). Examination of the DNA by electron microscopy confirmed that the bands seen in the agarose gels represented circular DNA molecules. The presence of plasmids was demonstrated in four additional strains. Three plasmids were carried in a strain of serovar 2, and single plasmids were detected in strains of serovars 4, 12, and 18. Plasmids were not detected in three strains of serovars 1, 3, and 5.
对鸟分枝杆菌-胞内分枝杆菌菌株进行了质粒检测。用环丝氨酸和氨苄青霉素处理对数生长期的培养物,以使细胞对去污剂裂解敏感,然后用十二烷基硫酸钠和氯化钠处理制备清亮裂解物。通过在含溴化乙锭的氯化铯梯度中进行平衡密度离心制备纯化的质粒DNA,并通过琼脂糖凝胶电泳进行分析。发现LR25菌株携带三种质粒,包括一种分子量大于10⁸的大质粒。通过电子显微镜检查DNA证实,琼脂糖凝胶中看到的条带代表环状DNA分子。在另外四个菌株中也证实了质粒的存在。血清型2的一个菌株携带三种质粒,血清型4、12和18的菌株中检测到单个质粒。在血清型1、3和5的三个菌株中未检测到质粒。