Watve Samit S, Bernardy Eryn E, Hammer Brian K
School of Biology, Georgia Institute of Technology, Atlanta, Georgia.
Curr Protoc Microbiol. 2014 Nov 3;35:6A.4.1-12. doi: 10.1002/9780471729259.mc06a04s35.
Many bacteria can become naturally competent to take up extracellular DNA across their outer and inner membranes by a dedicated competence apparatus. Whereas some studies show that the DNA delivered to the cytoplasm may be used for genome repair or for nutrition, it can also be recombined onto the chromosome by homologous recombination: a process called natural transformation. Along with conjugation and transduction, natural transformation represents a mechanism for horizontal transfer of genetic material, e.g., antibiotic resistance genes, which can confer new beneficial characteristics onto the recipient bacteria. Described here are protocols for quantifying the frequency of transformation for the human pathogen Vibrio cholerae, one of several Vibrio species recently shown to be capable of natural transformation.
许多细菌可通过专门的感受态装置自然获得摄取穿过其外膜和内膜的细胞外DNA的能力。虽然一些研究表明,递送至细胞质的DNA可用于基因组修复或营养,但它也可通过同源重组整合到染色体上:这一过程称为自然转化。与接合和转导一起,自然转化代表了一种遗传物质水平转移的机制,例如抗生素抗性基因,它可以赋予受体细菌新的有益特性。本文描述了定量人类病原体霍乱弧菌转化频率的实验方案,霍乱弧菌是最近被证明能够进行自然转化的几种弧菌之一。