Magrini V, Creighton C, White D, Hartzell P L, Youderian P
Department of Microbiology, Molecular Biology and Biochemistry, University of Idaho, Moscow, Idaho 83844-3052, USA.
J Bacteriol. 1998 Dec;180(24):6757-60. doi: 10.1128/JB.180.24.6757-6760.1998.
Plasmids with the aadA gene from plasmid R100, which confers resistance to the aminoglycosides spectinomycin and streptomycin in Escherchia coli, can be introduced into wild-type Myxococcus xanthus, strain DK1622, by electroporation. Recombinant M. xanthus strains with integrated plasmids carrying the aadA gene acquire resistance to high levels of these antibiotics. Selection for aadA in M. xanthus can be carried out independently of, or simultaneously with, selection for resistance to kanamycin. The kinds and frequencies of recombination events observed between integrative plasmids with aadA and the M. xanthus chromosome are similar to those observed after the transformation of yeast. Cleavage of integrative plasmid DNA at a site adjacent to a region of homology between the plasmid and the M. xanthus genome favors the targeted disruption of M. xanthus genes by allele replacement.
携带来自质粒R100的aadA基因的质粒，该基因赋予大肠杆菌对氨基糖苷类壮观霉素和链霉素的抗性，可通过电穿孔导入野生型黄色粘球菌DK1622菌株。携带aadA基因的整合质粒的重组黄色粘球菌菌株获得了对这些抗生素的高水平抗性。在黄色粘球菌中对aadA的选择可以独立于或与对卡那霉素抗性的选择同时进行。在携带aadA的整合质粒与黄色粘球菌染色体之间观察到的重组事件的种类和频率与酵母转化后观察到的相似。在与质粒和黄色粘球菌基因组之间的同源区域相邻的位点切割整合质粒DNA有利于通过等位基因置换对黄色粘球菌基因进行靶向破坏。
