All-in-one construct for genome engineering using Cre-lox technology
Martínez Mariscal, Ana M. (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Gonzalez-Gonzalez, Luis 1987- (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Querol Murillo, Enrique (Universitat Autònoma de Barcelona. Departament de Bioquímica i de Biologia Molecular)
Piñol Ribas, Jaume (Universitat Autònoma de Barcelona. Institut de Biotecnologia i de Biomedicina "Vicent Villar Palasí")

Date:	2016
Abstract:	Mycoplasma genitalium is an appealing model of a minimal cell and synthetic biology study, and it was one of the first organisms whose genome was fully sequenced and chemically synthesized. Despite its usefulness as a model organism, many genetic tools well established for other microorganisms are not currently available in mycoplasmas. We have developed several vectors to adapt the Cre-lox technology for genome engineering in M. genitalium, providing an all-in-one construct that could be also useful to obtain unmarked genetic modifications in many other slow growing microorganisms. This construct contains a modified promoter sequence based in TetR system that exhibits an enhanced control on Cre recombinase expression, virtually abolishing the presence of this recombinase in the absence of inducer. This allows to introduce the Cre recombinase gene and the desired genetic modification in a single transformation step. In addition, this inducible promoter may be a very promising tool for a wide range of molecular applications.
Grants:	Ministerio de Economía y Competitividad BIO2013-4870R Ministerio de Economía y Competitividad BIO2013-50176EXP Agència de Gestió d'Ajuts Universitaris i de Recerca FI-DGR 2014 Agència de Gestió d'Ajuts Universitaris i de Recerca FI-DGR 2012
Rights:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, sempre que no sigui amb finalitats comercials, i sempre que es reconegui l'autoria de l'obra original.
Language:	Anglès
Document:	Article ; recerca ; Versió publicada
Subject:	Cre-lox technology ; Mycoplasmas ; Genome engineering ; TetR-TetO inducible promoters
Published in:	DNA research, Vol. 23, issue 3 (June 2016) , p. 263-270, ISSN 1756-1663

DOI: 10.1093/dnares/dsw015
PMID: 27084897

8 p, 416.7 KB

The record appears in these collections:
Research literature > UAB research groups literature > Research Centres and Groups (research output) > Health sciences and biosciences > Institut de Biotecnologia i de Biomedicina (IBB)
Articles > Research articles
Articles > Published articles