Mu transpososome activity-profiling yields hyperactive MuA variants for highly efficient genetic and genome engineering.

ABSTRACT: The phage Mu DNA transposition system provides a versatile species non-specific tool for molecular biology, genetic engineering and genome modification applications. Mu transposition is catalyzed by MuA transposase, with DNA cleavage and integration reactions ultimately attaching the transposon DNA to target DNA. To improve the activity of the Mu DNA transposition machinery, we mutagenized MuA protein and screened for hyperactivity-causing substitutions using an in vivo assay. The individual activity-enhancing substitutions were mapped onto the MuA-DNA complex structure, containing a tetramer of MuA transposase, two Mu end segments and a target DNA. This analysis, combined with the varying effect of the mutations in different assays, implied that the mutations exert their effects in severa

SUBMITTER: Rasila TS 

PROVIDER: S-EPMC5961161 | biostudies-literature | 2018 May

REPOSITORIES: biostudies-literature