Creating bacterial strains from genomes that have been cloned and engineered in yeast.
Science, 2009/9/25;325(5948):1693-6.
Lartigue C[1], Vashee S, Algire MA, Chuang RY, Benders GA, Ma L, Noskov VN, Denisova EA, Gibson DG, Assad-Garcia N, Alperovich N, Thomas DW, Merryman C, Hutchison CA 3rd, Smith HO, Venter JC, Glass JI
Affiliations
PMID: 19696314DOI: 10.1126/science.1173759
Impact factor: 63.714
Abstract
We recently reported the chemical synthesis, assembly, and cloning of a bacterial genome in yeast. To produce a synthetic cell, the genome must be transferred from yeast to a receptive cytoplasm. Here we describe methods to accomplish this. We cloned a Mycoplasma mycoides genome as a yeast centromeric plasmid and then transplanted it into Mycoplasma capricolum to produce a viable M. mycoides cell. While in yeast, the genome was altered by using yeast genetic systems and then transplanted to produce a new strain of M. mycoides. These methods allow the construction of strains that could not be produced with genetic tools available for this bacterium.
MeSH terms
Centromere; Cloning, Molecular; DNA Methylation; DNA Restriction Enzymes; Deoxyribonucleases, Type III Site-Specific; Gene Transfer Techniques; Genetic Engineering; Genome, Bacterial; Mycoplasma capricolum; Mycoplasma mycoides; Plasmids; Saccharomyces cerevisiae; Sequence Analysis, DNA; Sequence Deletion; Transformation, Bacterial
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