An att site-based recombination reporter system for genome engineering and synthetic DNA assembly - Archive ouverte HAL Access content directly
Journal Articles BMC Biotechnology Year : 2017

An att site-based recombination reporter system for genome engineering and synthetic DNA assembly

(1) , (1) , (1) , (1)
1

Abstract

BACKGROUND: Direct manipulation of the genome is a widespread technique for genetic studies and synthetic biology applications. The tyrosine and serine site-specific recombination systems of bacteriophages HK022 and ΦC31 are widely used for stable directional exchange and relocation of DNA sequences, making them valuable tools in these contexts. We have developed site-specific recombination tools that allow the direct selection of recombination events by embedding the attB site from each system within the β-lactamase resistance coding sequence (bla). RESULTS: The HK and ΦC31 tools were developed by placing the attB sites from each system into the signal peptide cleavage site coding sequence of bla. All possible open reading frames (ORFs) were inserted and tested for recombination efficiency and bla activity. Efficient recombination was observed for all tested ORFs (3 for HK, 6 for ΦC31) as shown through a cointegrate formation assay. The bla gene with the embedded attB site was functional for eight of the nine constructs tested. CONCLUSIONS: The HK/ΦC31 att-bla system offers a simple way to directly select recombination events, thus enhancing the use of site-specific recombination systems for carrying out precise, large-scale DNA manipulation, and adding useful tools to the genetics toolbox. We further show the power and flexibility of bla to be used as a reporter for recombination.
Fichier principal
Vignette du fichier
BMC Biotechnol_Jul 2017.pdf (755.11 Ko) Télécharger le fichier
Origin : Publication funded by an institution
Loading...

Dates and versions

pasteur-01688696 , version 1 (19-01-2018)

Licence

Attribution - CC BY 4.0

Identifiers

Cite

Michael Bland, Magaly Ducos-Galand, Marie-Eve Val, Didier Mazel. An att site-based recombination reporter system for genome engineering and synthetic DNA assembly. BMC Biotechnology, 2017, 17 (1), pp.62. ⟨10.1186/s12896-017-0382-1⟩. ⟨pasteur-01688696⟩

Collections

PASTEUR CNRS ANR
31 View
166 Download

Altmetric

Share

Gmail Facebook Twitter LinkedIn More