Selection-free markerless genome manipulations in the polyploid bacterium Thermus thermophilus.

ABSTRACT: A genome manipulation approach based on double-crossover homologous recombination was developed in the polyploid model organism Thermus thermophilus HB27 without the use of any selectable marker. The method was established and optimized by targeting the megaplasmid-encoded β-glucosidase gene bgl. When linear and supercoiled forms of marker-free suicide vector were used for transformations, the frequencies of obtaining apparent Bgl^- mutant were 10^- 5 and 10^- 3, respectively; while the frequency could reach 10^- 2 when transformation with concatemer form of the same vector. All randomly selected Bgl^- colonies from the transformations were found to be true bgl knockout mutants. Thus, markerless gene deletion mutants co

SUBMITTER: Li H 

PROVIDER: S-EPMC6431304 | biostudies-literature | 2019 Apr

REPOSITORIES: biostudies-literature