Deletion of an essential gene in Escherichia coli by site-specific recombination with linear DNA fragments

作者：Maria Jasin, PAUL R. SCHIMMEL · 发表于：Journal of Bacteriology · 年份：1984 · DOI：10.1128/jb.159.2.783-786.1984 · 被引用次数：148 · 研究领域：Bacterial Genetics and Biotechnology、DNA Repair Mechanisms、RNA and protein synthesis mechanisms

Deletion of an essential gene in Escherichia coli was accomplished by transformation of linear DNA fragments that have a Kanr gene segment flanked by sequences homologous to closely spaced regions on the E. coli chromosome. Selection for a double crossover within homologous sequences can effectively delete an entire gene. Cell viability is maintained by provision of the essential gene on a plasmid with a temperature-sensitive replicon, resulting in cells which have a temperature-sensitive phenotype.