Escherichia coli vectors having stringently repressible replication origins allow a streamlining of Crispr/Cas9 gene editing.

ABSTRACT: Readily curable plasmids facilitate the construction of plasmid-free bacterial strains after the plasmid encoded genes are no longer needed. The most popular of these plasmids features a temperature-sensitive (Ts) pSC101 origin of replication which can readily revert during usage and cannot be used to construct Ts mutations in essential genes. Plasmid pAM34 which contains an IPTG-dependent origin of replication largely overcomes this issue but is limited by carrying the most commonly utilized antibiotic selection and replication origin. This study describes the construction of an expanded series of plasmid vectors having replication origins of p15a, RSF1030 or RSF1031 that like pAM34 have IPTG-dependent replication. Surprisingly, these plasmids can be cured in fewer generations than pAM34.

SUBMITTER: Srinivas S 

PROVIDER: S-EPMC7260698 | biostudies-literature | 2019 May

REPOSITORIES: biostudies-literature