Wang Y, Taylor D E
Department of Microbiology, University of Alberta, Edmonton, Canada.
J Bacteriol. 1990 Feb;172(2):949-55. doi: 10.1128/jb.172.2.949-955.1990.
Growing cells of Campylobacter coli and C. jejuni were naturally transformed by naked DNA without the requirement for any special treatment. Transformation frequencies for homologous chromosomal DNA were approximately 10(-3) transformants per recipient cell in C. coli and 10(-4) in C. jejuni. Maximum competence was found in the early log phase of growth. Campylobacters preferentially took up their own DNA in comparison with Escherichia coli chromosomal DNA, which was taken up very poorly. Three new Campylobacter spp.-to-E. coli shuttle plasmids, which contained additional cloning sites and selectable markers, were constructed from the shuttle vector pILL550A. These plasmid DNAs were taken up by campylobacters much less efficiently than was homologous chromosomal DNA, and transformation into plasmid-free cells was very rare. However, with the use of recipients containing a homologous plasmid, approximately 10(-4) transformants per cell were obtained. The tetM determinant, originally obtained from Streptococcus spp. and not heretofore reported in Campylobacter spp., was isolated from an E. coli plasmid and was introduced, selecting for tetracycline resistance, by natural transformation into C. coli.
空肠弯曲杆菌和结肠弯曲杆菌的生长细胞可被裸露的DNA自然转化,无需任何特殊处理。对于同源染色体DNA,结肠弯曲杆菌的转化频率约为每个受体细胞10^(-3)个转化子,空肠弯曲杆菌为10^(-4)个。在生长的对数早期发现最大感受态。与摄取很差的大肠杆菌染色体DNA相比,弯曲杆菌优先摄取自身的DNA。从穿梭载体pILL550A构建了三种新的从弯曲杆菌属到大肠杆菌的穿梭质粒,它们含有额外的克隆位点和选择标记。这些质粒DNA被弯曲杆菌摄取的效率远低于同源染色体DNA,转化为无质粒细胞的情况非常罕见。然而,使用含有同源质粒的受体,每个细胞可获得约10^(-4)个转化子。tetM决定簇最初从链球菌属获得,此前在弯曲杆菌属中未见报道,从大肠杆菌质粒中分离出来,并通过自然转化导入结肠弯曲杆菌,筛选四环素抗性。