重组工程
溶解循环
支原体
噬菌体
生物
DNA
遗传学
同源重组
基因
病毒
大肠杆菌
医学
病理
结核分枝杆菌
肺结核
作者
Laura J. Marinelli,Mariana Piuri,Graham F. Hatfull
出处
期刊:Methods in molecular biology
日期:2018-12-21
卷期号:: 69-80
被引量:15
标识
DOI:10.1007/978-1-4939-8940-9_6
摘要
We describe a recombineering-based method for the genetic manipulation of lytically replicating bacteriophages, focusing on mycobacteriophages. The approach utilizes recombineering-proficient strains of Mycobacterium smegmatis and 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 in M. smegmatis recombineering strains, allows for the efficient and rapid generation of bacteriophage mutants.
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