Development of a Suite of Tools for Genome Editing in <i>Parageobacillus thermoglucosidasius</i> and Their Use to Identify the Potential of a Native Plasmid in the Generation of Stable Engineered Strains
Matthew S. H. Lau, Lili Sheng, Ying Zhang, Nigel P. Minton
Abstract
, pNCI001 and pNCI002, either singly or in combination, had no discernible effects on the overall phenotypic characteristics of the organism. Through the CRISPR/Cas9-mediated insertion of the gene encoding a novel fluorescent reporter, eCGP123, we showed that pNCI001 exhibited a high degree of segregational stability. As the relatively higher copy number of pNCI001 led to higher levels of eCGP123 expression than when the same gene was integrated into the chromosome, we propose that pNCI001 represents the preferred option for the integration of metabolic operons when stable commercial strains are required.
Topics & Concepts
Genome editingCas9CRISPRPlasmidBiologyComputational biologyGenome engineeringChassisSynthetic biologyGeneGenomeRecombineeringGeneticsEngineeringStructural engineeringCRISPR and Genetic EngineeringBacterial Genetics and BiotechnologyBacteriophages and microbial interactions