Natural Cotransformation and Multiplex Genome Editing by Natural Transformation (MuGENT) of Vibrio cholerae
Natural Cotransformation and Multiplex Genome Editing by Natural Transformation (MuGENT) of Vibrio cholerae
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DOI：
10.1007/978-1-4939-8685-9_6
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发表时间：
2018-01-01
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影响因子：
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通讯作者：
Dalia, Ankur B.
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作者：
Dalia, Ankur B.
产生突变菌株是微生物遗传学的重要组成部分。天然遗传转化是一种吸收和整合外源DNA的过程，由不同的微生物物种共享，并可用于制备突变菌株。规范上，该方法已用于产生单个突变体，并依次产生具有多个突变的菌株。最近，我们描述了一种通过自然转化进行多重基因组编辑的方法（MuGENT），该方法允许在单个步骤中产生具有多个无瘢痕突变的菌株。在这里，我们提供了一个用于霍乱病原体霍乱弧菌的诱变方法的详细描述，特别强调通过MuGENT诱变。
Generating mutant strains is an essential component of microbial genetics. Natural genetic transformation, a process for the uptake and integration of foreign DNA, is shared by diverse microbial species and can be exploited for making mutant strains. Canonically, this process has been used to generate single mutants and sequentially to generate strains with multiple mutations. Recently, we have described a method for multiplex genome editing by natural transformation (MuGENT), which allows the generation of strains with multiple scarless mutations in a single step. Here, we provide a detailed description of the methods used for mutagenesis of the cholera pathogen Vibrio cholerae with a particular emphasis on mutagenesis via MuGENT.