A targetron system for gene targeting in thermophiles and its application in Clostridium thermocellum.

ABSTRACT:

Background

Targetrons are gene targeting vectors derived from mobile group II introns. They consist of an autocatalytic intron RNA (a "ribozyme") and an intron-encoded reverse transcriptase, which use their combined activities to achieve highly efficient site-specific DNA integration with readily programmable DNA target specificity.

Methodology/principal findings

Here, we used a mobile group II intron from the thermophilic cyanobacterium Thermosynechococcus elongatus to construct a thermotargetron for gene targeting in thermophiles. After determining its DNA targeting rules by intron mobility assays in Escherichia coli at elevated temperatures, we used this thermotargetron in Clostridium thermocellum, a thermophile employed in biofuels production, to disrupt six different ch

SUBMITTER: Mohr G 

PROVIDER: S-EPMC3706431 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature