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.
authors.affiliation: Institute of Biological, Environmental and Rural Sciences, Edward Llwyd Building, Aberystwyth University, Aberystwyth SY23 3FG, UK.
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Zampini
authors.affiliation: Institute of Biological, Environmental and Rural Sciences, Edward Llwyd Building, Aberystwyth University, Aberystwyth SY23 3FG, UK.
authors.forename: Pauline Rees
authors.initials: PR
authors.lastname: Stevens
authors.affiliation: Institute of Biological, Environmental and Rural Sciences, Edward Llwyd Building, Aberystwyth University, Aberystwyth SY23 3FG, UK.
authors.forename: Justin A
authors.initials: JA
authors.lastname: Pachebat
authors.affiliation: Institute of Biological, Environmental and Rural Sciences, Edward Llwyd Building, Aberystwyth University, Aberystwyth SY23 3FG, UK.
authors.forename: Alison
authors.initials: A
authors.lastname: Kingston-Smith
authors.affiliation: Institute of Biological, Environmental and Rural Sciences, Edward Llwyd Building, Aberystwyth University, Aberystwyth SY23 3FG, UK.
authors.forename: Luis A J
authors.initials: LA
authors.lastname: Mur
authors.affiliation: Faculty of Life Sciences, University of Manchester, Manchester M13 9PL, UK.
authors.forename: Finbarr
authors.initials: F
authors.lastname: Hayes
mesh_terms: D000818:Animals; D004247:DNA / Q000138:chemical synthesis* / Q000235:genetics; D024881:Drug Resistance, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000235:genetics; D050982:GATA1 Transcription Factor / Q000235:genetics; D005798:Genes, Bacterial / Q000235:genetics; D005813:Genes, Synthetic; D005818:Genetic Engineering / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000235:genetics; D037402:Hydrozoa / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D000198:Spectinomycin / Q000494:pharmacology; D058615:Synthetic Biology / Q000379:methods*
chemical_list: D029968:Escherichia coli Proteins; D050982:GATA1 Transcription Factor; C485192:green fluorescent protein, Aequorea victoria; D049452:Green Fluorescent Proteins; D004247:DNA; D000198:Spectinomycin