RapGene: a fast and accurate strategy for synthetic gene assembly in Escherichia coli

Massimiliano Zampini; Pauline Rees Stevens; Justin Pachebat; Alison Kingston-Smith; Luis Mur; Finbarr Hayes

doi:10.1038/srep11302

RapGene: a fast and accurate strategy for synthetic gene assembly in Escherichia coli

Massimiliano Zampini, Pauline Rees Stevens, Justin Pachebat, Alison Kingston-Smith, Luis Mur, Finbarr Hayes

University of Manchester

Research output: Contribution to journal › Article › peer-review

7 Citations (Scopus)

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Abstract

The ability to assemble DNA sequences de novo through efficient and powerful DNA fabrication methods is one of the foundational technologies of synthetic biology. Gene synthesis, in particular, has been considered the main driver for the emergence of this new scientific discipline. Here we describe RapGene, a rapid gene assembly technique which was successfully tested for the synthesis and cloning of both prokaryotic and eukaryotic genes through a ligation independent approach. The method developed in this study is a complete bacterial gene synthesis platform for the quick, accurate and cost effective fabrication and cloning of gene-length sequences that employ the widely used host Escherichia coli.

Original language	English
Article number	11302
Journal	Scientific Reports
Volume	5
DOIs	https://doi.org/10.1038/srep11302
Publication status	Published - 11 Jun 2015

Keywords

genetic engineering
applied microbiology
DNA recombination
metabolic engineering
Synthetic Biology/methods
Genes, Synthetic
DNA/chemical synthesis
Escherichia coli/genetics
Hydrozoa/genetics
Genes, Bacterial/genetics
GATA1 Transcription Factor/genetics
Animals
Polymerase Chain Reaction/methods
Spectinomycin/pharmacology
Genetic Engineering/methods
Drug Resistance, Bacterial/genetics
Green Fluorescent Proteins/genetics
Escherichia coli Proteins/genetics

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10.1038/srep11302

RapGene: a fast and accurate strategy for synthetic gene assembly in Escherichia coliFinal published version, 836 KB
Supplementary InformationData, 259 KBLicence: CC BY

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title = "RapGene: a fast and accurate strategy for synthetic gene assembly in Escherichia coli",

abstract = "The ability to assemble DNA sequences de novo through efficient and powerful DNA fabrication methods is one of the foundational technologies of synthetic biology. Gene synthesis, in particular, has been considered the main driver for the emergence of this new scientific discipline. Here we describe RapGene, a rapid gene assembly technique which was successfully tested for the synthesis and cloning of both prokaryotic and eukaryotic genes through a ligation independent approach. The method developed in this study is a complete bacterial gene synthesis platform for the quick, accurate and cost effective fabrication and cloning of gene-length sequences that employ the widely used host Escherichia coli.",

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author = "Massimiliano Zampini and {Rees Stevens}, Pauline and Justin Pachebat and Alison Kingston-Smith and Luis Mur and Finbarr Hayes",

note = "Sponsorship: BBSRC RONO: BBS/E/W/10964A01; BB/G003114/1 Funding Information: We thank Miss Ainhoa Dafis-Sagarmendi for helping with PCR. This work was supported by the Biotechnology and Biological Sciences Research Council [grant numbers BBS/E/W/10964A to A.K-S, M.Z. & P.R.S, BB/G003114/1 to F.H.]. Funding for open access charge: Biotechnology and Biological Sciences Research Council.",

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doi = "10.1038/srep11302",

language = "English",

volume = "5",

journal = "Scientific Reports",

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AU - Pachebat, Justin

AU - Kingston-Smith, Alison

AU - Mur, Luis

AU - Hayes, Finbarr

N1 - Sponsorship: BBSRC RONO: BBS/E/W/10964A01; BB/G003114/1 Funding Information: We thank Miss Ainhoa Dafis-Sagarmendi for helping with PCR. This work was supported by the Biotechnology and Biological Sciences Research Council [grant numbers BBS/E/W/10964A to A.K-S, M.Z. & P.R.S, BB/G003114/1 to F.H.]. Funding for open access charge: Biotechnology and Biological Sciences Research Council.

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KW - genetic engineering

KW - applied microbiology

KW - DNA recombination

KW - metabolic engineering

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KW - Escherichia coli/genetics

KW - Hydrozoa/genetics

KW - Genes, Bacterial/genetics

KW - GATA1 Transcription Factor/genetics

KW - Animals

KW - Polymerase Chain Reaction/methods

KW - Spectinomycin/pharmacology

KW - Genetic Engineering/methods

KW - Drug Resistance, Bacterial/genetics

KW - Green Fluorescent Proteins/genetics

KW - Escherichia coli Proteins/genetics

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SN - 2045-2322

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RapGene: a fast and accurate strategy for synthetic gene assembly in Escherichia coli

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Alison Kingston-Smith

RSB : Rumen Systems Biology

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RapGene: a fast and accurate strategy for synthetic gene assembly in Escherichia coli

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