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PMID: 19073939 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

One-step assembly in yeast of 25 overlapping DNA fragments to form a complete synthetic Mycoplasma genitalium genome.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 105 ·No. 51 ·2008-12-23 ·Pages 20404-9

Gibson DG, Benders GA, Axelrod KC, Zaveri J, Algire MA, Moodie M, Montague MG, Venter JC, Smith HO, Hutchison CA

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.

MeSH Terms
Cloning, Molecular/methods DNA/biosynthesis Genome, Bacterial/genetics Mycoplasma genitalium/genetics Oligodeoxyribonucleotides/genetics,metabolism Recombination, Genetic Yeasts/genetics
Chemicals
Oligodeoxyribonucleotides DNA
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Gibson Daniel G
The J. Craig Venter Institute, Synthetic Biology Group, Rockville, MD 20850, USA. dgibson@jcvi.org
Benders Gwynedd A
Axelrod Kevin C
Zaveri Jayshree
Algire Mikkel A
Moodie Monzia
Montague Michael G
Venter J Craig
Smith Hamilton O
Hutchison Clyde A
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
1091-6490
DOI
10.1073/pnas.0811011106
Email
dgibson@jcvi.org
Published
2008-12-23
Epub
2008-00-10
Pages
20404-9
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC2600582
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.0811011106 PMC Full Text View journal details
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