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肺炎链球菌的电转化和自然转化:重组所需的DNA加工过程

Electrotransformation and natural transformation of Streptococcus pneumoniae: requirement of DNA processing for recombination.

作者信息

LefranGois Jacques, Samrakandi M Mustapha, Sicard A Michel

机构信息

Laboratoire de Microbiologie et de GCnCtique MolCculaire du CNRS and UniversitC Paul Sabatier, 118 route de Narbonnei 31062 Toulouse Cedex, France.

出版信息

Microbiology (Reading). 1998 Nov;144 ( Pt 11):3061-3068. doi: 10.1099/00221287-144-11-3061.

Abstract

Electrotransformation has been used as a tool to introduce genes carried on replicative vectors in hundreds of bacterial species. In this study, the technique was used to try to obtain recombination of markers in the chromosome of the naturally transformable bacterium Streptococcus pneumoniae. Recombination was not observed even using naturally competent cultures. Both chromosomal and cloned DNA, denatured or native, were without effect. These results suggest that it is not sufficient to introduce DNA into the cell to obtain recombinants in this bacterium. The integration of markers into the chromosome in naturally competent cells must require DNA processing during entry. Electrotransformation of replicating plasmids is recA-independent but can be facilitated by a recA-dependent process. This facilitation required the induction of the natural competence machinery, probably involving partial homologous pairing.

摘要

电转化已被用作一种工具,用于将携带在复制型载体上的基因导入数百种细菌中。在本研究中,该技术被用于尝试在天然可转化细菌肺炎链球菌的染色体中获得标记重组。即使使用天然感受态培养物,也未观察到重组。无论是染色体DNA还是克隆DNA,变性的或天然的,都没有效果。这些结果表明,将DNA导入细胞不足以在这种细菌中获得重组体。在天然感受态细胞中,标记整合到染色体中必定需要在DNA进入过程中进行加工。复制型质粒的电转化不依赖recA,但可以由一个依赖recA的过程来促进。这种促进需要诱导天然感受态机制,可能涉及部分同源配对。

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