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A set of vectors for introduction of antibiotic resistance genes by in vitro Cre-mediated recombination

BACKGROUND: Introduction of new antibiotic resistance genes in the plasmids of interest is a frequent task in molecular cloning practice. Classical approaches involving digestion with restriction endonucleases and ligation are time-consuming. FINDINGS: We have created a set of insertion vectors (pIN...

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Autores principales: Dmitriev, Petr V, Vassetzky, Yegor S
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2654900/
https://www.ncbi.nlm.nih.gov/pubmed/19105832
http://dx.doi.org/10.1186/1756-0500-1-135
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author Dmitriev, Petr V
Vassetzky, Yegor S
author_facet Dmitriev, Petr V
Vassetzky, Yegor S
author_sort Dmitriev, Petr V
collection PubMed
description BACKGROUND: Introduction of new antibiotic resistance genes in the plasmids of interest is a frequent task in molecular cloning practice. Classical approaches involving digestion with restriction endonucleases and ligation are time-consuming. FINDINGS: We have created a set of insertion vectors (pINS) carrying genes that provide resistance to various antibiotics (puromycin, blasticidin and G418) and containing a loxP site. Each vector (pINS-Puro, pINS-Blast or pINS-Neo) contains either a chloramphenicol or a kanamycin resistance gene and is unable to replicate in most E. coli strains as it contains a conditional R6Kγ replication origin. Introduction of the antibiotic resistance genes into the vector of interest is achieved by Cre-mediated recombination between the replication-incompetent pINS and a replication-competent target vector. The recombination mix is then transformed into E. coli and selected by the resistance marker (kanamycin or chloramphenicol) present in pINS, which allows to recover the recombinant plasmids with 100% efficiency. CONCLUSION: Here we propose a simple strategy that allows to introduce various antibiotic-resistance genes into any plasmid containing a replication origin, an ampicillin resistance gene and a loxP site.
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spelling pubmed-26549002009-03-13 A set of vectors for introduction of antibiotic resistance genes by in vitro Cre-mediated recombination Dmitriev, Petr V Vassetzky, Yegor S BMC Res Notes Technical Note BACKGROUND: Introduction of new antibiotic resistance genes in the plasmids of interest is a frequent task in molecular cloning practice. Classical approaches involving digestion with restriction endonucleases and ligation are time-consuming. FINDINGS: We have created a set of insertion vectors (pINS) carrying genes that provide resistance to various antibiotics (puromycin, blasticidin and G418) and containing a loxP site. Each vector (pINS-Puro, pINS-Blast or pINS-Neo) contains either a chloramphenicol or a kanamycin resistance gene and is unable to replicate in most E. coli strains as it contains a conditional R6Kγ replication origin. Introduction of the antibiotic resistance genes into the vector of interest is achieved by Cre-mediated recombination between the replication-incompetent pINS and a replication-competent target vector. The recombination mix is then transformed into E. coli and selected by the resistance marker (kanamycin or chloramphenicol) present in pINS, which allows to recover the recombinant plasmids with 100% efficiency. CONCLUSION: Here we propose a simple strategy that allows to introduce various antibiotic-resistance genes into any plasmid containing a replication origin, an ampicillin resistance gene and a loxP site. BioMed Central 2008-12-23 /pmc/articles/PMC2654900/ /pubmed/19105832 http://dx.doi.org/10.1186/1756-0500-1-135 Text en Copyright © 2008 Vassetzky et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Technical Note
Dmitriev, Petr V
Vassetzky, Yegor S
A set of vectors for introduction of antibiotic resistance genes by in vitro Cre-mediated recombination
title A set of vectors for introduction of antibiotic resistance genes by in vitro Cre-mediated recombination
title_full A set of vectors for introduction of antibiotic resistance genes by in vitro Cre-mediated recombination
title_fullStr A set of vectors for introduction of antibiotic resistance genes by in vitro Cre-mediated recombination
title_full_unstemmed A set of vectors for introduction of antibiotic resistance genes by in vitro Cre-mediated recombination
title_short A set of vectors for introduction of antibiotic resistance genes by in vitro Cre-mediated recombination
title_sort set of vectors for introduction of antibiotic resistance genes by in vitro cre-mediated recombination
topic Technical Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2654900/
https://www.ncbi.nlm.nih.gov/pubmed/19105832
http://dx.doi.org/10.1186/1756-0500-1-135
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