Department of Oral Biology, College of Dentistry, The University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA.
FEMS Microbiol Lett. 2011 Sep;322(2):138-44. doi: 10.1111/j.1574-6968.2011.02344.x. Epub 2011 Jul 18.
Veillonellae are one of the most prevalent and predominant microorganisms in both the supra- and subgingival plaques of the human oral cavity. Veillonellae's mutualistic relationships with the early, middle, and late colonizers of the oral cavity make them an important component of oral biofilm ecology. Unlike other ubiquitous early colonizers in the oral cavity, surprisingly little is known about Veillonella biology due to our lack of ability to genetically transform this group of bacteria. The objective of this study was to test the transformability of veillonellae. Using Veillonella parvula strain PK1910, we first obtained spontaneous mutations conferring streptomycin resistance. These mutations all carry a K43N substitution in the RpsL protein. Using the mutated rpsL gene as a selection marker, a variety of conditions were tested and optimized for electroporation. With the optimized protocol, we were able to introduce the first targeted mutation into the chromosome of V. parvula PK1910. Although more studies are needed to develop a robust genetic manipulation system in veillonellae, our results demonstrated, for the first time, that V. parvula is transformable, at least for strain PK1910.
韦荣氏球菌是人类口腔的龈上和龈下菌斑中最普遍和主要的微生物之一。韦荣氏球菌与口腔早期、中期和晚期定植者的共生关系使它们成为口腔生物膜生态学的重要组成部分。与口腔中其他普遍存在的早期定植者不同,由于我们缺乏对这群细菌进行基因转化的能力,因此对韦荣氏球菌的生物学特性知之甚少。本研究旨在测试韦荣氏球菌的可转化性。使用韦荣氏球菌 PK1910 株,我们首先获得了赋予链霉素抗性的自发突变。这些突变均在 RpsL 蛋白中携带 K43N 取代。使用突变的 rpsL 基因作为选择标记,我们测试并优化了各种电穿孔条件。使用优化的方案,我们能够将第一个靶向突变引入 V. parvula PK1910 的染色体中。尽管需要进一步的研究来开发韦荣氏球菌中稳健的遗传操作系统,但我们的结果首次表明,至少对于 PK1910 株,韦荣氏球菌是可转化的。