TY - JOUR
T1 - RapGene
T2 - a fast and accurate strategy for synthetic gene assembly in Escherichia coli
AU - Zampini, Massimiliano
AU - Rees Stevens, Pauline
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.
PY - 2015/6/11
Y1 - 2015/6/11
N2 - 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.
AB - 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.
KW - genetic engineering
KW - applied microbiology
KW - DNA recombination
KW - metabolic engineering
KW - Synthetic Biology/methods
KW - Genes, Synthetic
KW - DNA/chemical synthesis
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
UR - http://www.scopus.com/inward/record.url?scp=84931287741&partnerID=8YFLogxK
U2 - 10.1038/srep11302
DO - 10.1038/srep11302
M3 - Article
C2 - 26062748
SN - 2045-2322
VL - 5
JO - Scientific Reports
JF - Scientific Reports
M1 - 11302
ER -
