Rescue of mutant fitness defects using in vitro reconstituted designer transposons in Mycoplasma mycoides.

ABSTRACT: With only hundreds of genes contained within their genomes, mycoplasmas have become model organisms for precise understanding of cellular processes, as well as platform organisms for predictable engineering of microbial functions for mission-critical applications. Despite the availability of "whole genome writing" in Mycoplasma mycoides, some traditional methods for genetic engineering are underdeveloped in mycoplasmas. Here we demonstrate two facile transposon-mediated approaches for introducing genes into the synthetic cell based on M. mycoides. The marker-less approach involves preparing a fragment containing only a small genomic region of interest with flanking transposase-binding sites, followed by in vitro transposase loading and introduction into the cells. The marker-driven approac

SUBMITTER: Karas BJ 

PROVIDER: S-EPMC4107850 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature