JCVI: Abstract
 
 
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Citation

Gibson, D. G., Benders, G. A., Axelrod, K. C., Zaveri, J., Algire, M. A., Moodie, M., Montague, M. G., Venter, J. C., Smith, H. O., Hutchison, C. A., 3rd

One-step Assembly In Yeast of 25 Overlapping DNA Fragments to Form a Complete Synthetic Mycoplasma genitalium Genome

Proc Natl Acad Sci U S A. 2008 Dec 23; 105(51): 20404-9.

PubMed Citation

Abstract

We previously reported assembly and cloning of the synthetic Mycoplasma genitalium JCVI-1.0 genome in the yeast Saccharomyces cerevisiae by recombination of six overlapping DNA fragments to produce a 592-kb circle. Here we extend this approach by demonstrating assembly of the synthetic genome from 25 overlapping fragments in a single step. The use of yeast recombination greatly simplifies the assembly of large DNA molecules from both synthetic and natural fragments.