清脆的
毒力
基因组编辑
质粒
突变
霍乱弧菌
生物
噬菌体
基因
基因组
遗传学
计算生物学
细菌
突变
大肠杆菌
出处
期刊:PubMed
日期:2023-04-28
标识
DOI:10.1101/pdb.prot108316
摘要
Tools for site-directed mutagenesis of virulent bacteriophages (phages; viruses of bacteria) have traditionally lagged those for bacteria, hindering their study. CRISPR gene editing represents a new and highly efficient method for editing virulent phage genomes. Here, I describe methods for using CRISPR gene editing for site-directed mutagenesis of ICP1, a virulent phage of Vibrio cholerae The first section outlines methods of constructing a plasmid for CRISPR editing of an ICP1 gene. The second section outlines methods of transferring the plasmid to an editing-competent strain of V. cholerae The third section outlines methods of selecting for and storing the edited phage.
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