Genetic Manipulation of Lytic Bacteriophages with BRED: Bacteriophage Recombineering of Electroporated DNA.
Genetic Manipulation of Lytic Bacteriophages with BRED: Bacteriophage Recombineering of Electroporated DNA.
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使用 BRED 对裂解性噬菌体进行基因操作：电穿孔 DNA 的噬菌体重组工程。
DOI：
10.1007/978-1-4939-8940-9_6
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Hatfull,GrahamF
中科院分区：
文献类型：
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作者：
Marinelli,LauraJ;Piuri,Mariana;Hatfull,GrahamF
我们描述了一种基于重组的方法，用于裂解复制噬菌体的遗传操作，重点是分枝杆菌噬菌体。该方法利用重组能力强的耻垢分枝杆菌菌株，并采用纯化的噬菌体基因组DNA和诱变底物的共转化策略，仅选择那些有能力摄取DNA的细胞。共转化方法结合M. smegmatisrecombinengineering菌株，允许有效和快速产生噬菌体突变体。
We describe a recombineering-based method for the genetic manipulation of lytically replicating bacteriophages, focusing on mycobacteriophages. The approach utilizes recombineering-proficient strains ofMycobacterium smegmatisand employs a cotransformation strategy with purified phage genomic DNA and a mutagenic substrate, which selects for only those cells that are competent to take up DNA. The cotransformation method, combined with the high rates of recombination obtained inM. smegmatisrecombineering strains, allows for the efficient and rapid generation of bacteriophage mutants.