Mahairas G G, Minion F C
Veterinary Medical Research Institute, College of Veterinary Medicine, Iowa State University, Ames 50011.
Plasmid. 1989 Jan;21(1):43-7. doi: 10.1016/0147-619x(89)90085-1.
The staphylococcal transposon Tn4001 was introduced into Mycoplasma pulmonis using an Escherichia coli-derived vector by polyethylene glycol-mediated transformation. Using a reaction mixture containing 10 micrograms plasmid DNA, 10 micrograms yeast tRNA, and 34-35% polyethylene glycol per 1 x 10(8) cells, Tn4001 could be introduced into M. pulmonis at a frequency of 5 x 10(-5) per colony forming unit. DNA-DNA hybridization studies illustrated that Tn4001 could occupy a diversity of insertion sites in the M. pulmonis chromosome. These data indicated that Tn4001 is a potentially useful tool for the introduction of mutations and for genetic studies in M. pulmonis.
利用大肠杆菌衍生载体，通过聚乙二醇介导的转化方法，将葡萄球菌转座子Tn4001导入肺支原体。每1×10⁸个细胞使用含有10微克质粒DNA、10微克酵母tRNA和34 - 35%聚乙二醇的反应混合物，Tn4001可以以每集落形成单位5×10⁻⁵的频率导入肺支原体。DNA - DNA杂交研究表明，Tn4001可占据肺支原体染色体中的多种插入位点。这些数据表明，Tn4001是在肺支原体中引入突变和进行遗传研究的潜在有用工具。
