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Originally published in Science Express on 20 August 2009
Science 25 September 2009:
Vol. 325. no. 5948, pp. 1693 - 1696
DOI: 10.1126/science.1173759
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Reports

Creating Bacterial Strains from Genomes That Have Been Cloned and Engineered in Yeast

Carole Lartigue,1 Sanjay Vashee,1,{dagger} Mikkel A. Algire,1 Ray-Yuan Chuang,1 Gwynedd A. Benders,2 Li Ma,1 Vladimir N. Noskov,1 Evgeniya A. Denisova,1 Daniel G. Gibson,1 Nacyra Assad-Garcia,1 Nina Alperovich,1 David W. Thomas,1,* Chuck Merryman,1 Clyde A. Hutchison, III,2 Hamilton O. Smith,2 J. Craig Venter,1,2 John I. Glass1

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.

1 The J. Craig Venter Institute, 9704 Medical Center Drive, Rockville, MD 20850, USA.
2 The J. Craig Venter Institute, 10355 Science Center Drive, San Diego, CA 92121, USA.

* Present address: Biotechnology Industry Organization (BIO), 1201 Maryland Avenue SW, Washington, DC 20024, USA.

{dagger} To whom correspondence should be addressed. E-mail: svashee{at}jcvi.org

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