A new large-DNA-fragment delivery system based on integrase activity from an integrative and conjugative element.

ABSTRACT: During the past few decades, numerous plasmid vectors have been developed for cloning, gene expression analysis, and genetic engineering. Cloning procedures typically rely on PCR amplification, DNA fragment restriction digestion, recovery, and ligation, but increasingly, procedures are being developed to assemble large synthetic DNAs. In this study, we developed a new gene delivery system using the integrase activity of an integrative and conjugative element (ICE). The advantage of the integrase-based delivery is that it can stably introduce a large DNA fragment (at least 75 kb) into one or more specific sites (the gene for glycine-accepting tRNA) on a target chromosome. Integrase recombination activity in Escherichia coli is kept low by using a synthetic hybrid promoter, which, however, i

SUBMITTER: Miyazaki R 

PROVIDER: S-EPMC3697514 | biostudies-literature | 2013 Jul

REPOSITORIES: biostudies-literature