Parallel in vivo DNA assembly by recombination: experimental demonstration and theoretical approaches

The development of synthetic biology requires rapid batch construction of large gene networks from combinations of smaller units. Despite the availability of computational predictions for well-characterized enzymes, the optimization of most synthetic biology projects requires combinational construct...

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Bibliographic Details
Published inPloS one Vol. 8; no. 2; p. e56854
Main Authors Shi, Zhenyu, Wedd, Anthony G, Gras, Sally L
Format Journal Article
LanguageEnglish
Published United States Public Library of Science 28.02.2013
Public Library of Science (PLoS)
Subjects
Assembly
Bioinformatics
Biology
Biotechnology
Cloning
Cloning, Molecular - methods
Computer applications
Construction
Construction materials
Deoxyribonucleic acid
Design
DNA
DNA - chemistry
DNA - metabolism
Enzymes
Gene Order
Genes
Genetic engineering
Genetic Vectors
Genomes
In vivo methods and tests
Metabolism
Methods
Optimization
Plasmids
Proteins
Recombination
Recombination, Genetic
Scars
Simulation
Synthetic biology
Victoria Australia
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Summary:The development of synthetic biology requires rapid batch construction of large gene networks from combinations of smaller units. Despite the availability of computational predictions for well-characterized enzymes, the optimization of most synthetic biology projects requires combinational constructions and tests. A new building-brick-style parallel DNA assembly framework for simple and flexible batch construction is presented here. It is based on robust recombination steps and allows a variety of DNA assembly techniques to be organized for complex constructions (with or without scars). The assembly of five DNA fragments into a host genome was performed as an experimental demonstration.
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Competing Interests: The authors have declared that no competing interests exist.
Helped revise the manuscript: SLG AGW. Conceived and designed the experiments: ZS. Performed the experiments: ZS. Analyzed the data: ZS. Contributed reagents/materials/analysis tools: ZS AGW. Wrote the paper: ZS.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0056854