Transposon mutagenesis of the extremely thermophilic bacterium Thermus thermophilus HB27.

ABSTRACT: Thermus thermophilus is an extremely thermophilic bacterium that grows between 50 and 80 °C and is an excellent model organism not only for understanding life at high temperature but also for its biotechnological and industrial applications. Multiple molecular capabilities are available including targeted gene inactivation and the use of shuttle plasmids that replicate in T. thermophilus and Escherichia coli; however, the ability to disrupt gene function randomly by transposon insertion has not been developed. Here we report a detailed method of transposon mutagenesis of T. thermophilus HB27 based on the EZ-Tn5 system from Epicentre Biotechnologies. We were able to generate insertion mutations throughout the chromosome by in vitro transposition and transformation with mutagenized genomic D

SUBMITTER: Carr JF 

PROVIDER: S-EPMC4272677 | biostudies-literature | 2015 Jan

REPOSITORIES: biostudies-literature