Cre-loxP recombination vectors for promoter studies

For promoter studies the cloning, subcloning and transfer to different plasmid vectors usually requires use of restriction enzymes and ligation reactions. One obstacle is the nucleotide polymorphisms of eukaryotic genomic DNA, which has the consequence that a sequence often differs from published se...

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Autores principales:	Pedersen,Nina, Tuxen Poulsen,Thomas, Skovgaard Poulsen,Hans
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2007
Materias:	cloning Cre recombinase plasmid reporter genes therapeutic genes
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582007000200013
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spelling	oai:scielo:S0717-345820070002000132007-10-29Cre-loxP recombination vectors for promoter studiesPedersen,NinaTuxen Poulsen,ThomasSkovgaard Poulsen,Hans cloning Cre recombinase plasmid reporter genes therapeutic genes For promoter studies the cloning, subcloning and transfer to different plasmid vectors usually requires use of restriction enzymes and ligation reactions. One obstacle is the nucleotide polymorphisms of eukaryotic genomic DNA, which has the consequence that a sequence often differs from published sequences. Therefore sequencing, rigorous restriction enzyme analysis or introduction of suitable sites has to be performed prior to cloning and subcloning. In addition, conventional methods using restriction enzymes, insert purifications and ligations is expensive and labour demanding. We have developed a fast, efficient and inexpensive Cre recombinase-loxP based method, which allows cloning of promoter regions and subcloning of these into a variety of vectors in a restriction enzyme independent manner. We here demonstrate that expression of a number of reporter genes and a therapeutic gene from both a viral and 2 mammalian promoters cloned by this recombinase method have activities comparable to conventionally cloned plasmidsinfo:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.10 n.2 20072007-04-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582007000200013en10.4067/S0717-34582007000200013
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	cloning Cre recombinase plasmid reporter genes therapeutic genes
spellingShingle	cloning Cre recombinase plasmid reporter genes therapeutic genes Pedersen,Nina Tuxen Poulsen,Thomas Skovgaard Poulsen,Hans Cre-loxP recombination vectors for promoter studies
description	For promoter studies the cloning, subcloning and transfer to different plasmid vectors usually requires use of restriction enzymes and ligation reactions. One obstacle is the nucleotide polymorphisms of eukaryotic genomic DNA, which has the consequence that a sequence often differs from published sequences. Therefore sequencing, rigorous restriction enzyme analysis or introduction of suitable sites has to be performed prior to cloning and subcloning. In addition, conventional methods using restriction enzymes, insert purifications and ligations is expensive and labour demanding. We have developed a fast, efficient and inexpensive Cre recombinase-loxP based method, which allows cloning of promoter regions and subcloning of these into a variety of vectors in a restriction enzyme independent manner. We here demonstrate that expression of a number of reporter genes and a therapeutic gene from both a viral and 2 mammalian promoters cloned by this recombinase method have activities comparable to conventionally cloned plasmids
author	Pedersen,Nina Tuxen Poulsen,Thomas Skovgaard Poulsen,Hans
author_facet	Pedersen,Nina Tuxen Poulsen,Thomas Skovgaard Poulsen,Hans
author_sort	Pedersen,Nina
title	Cre-loxP recombination vectors for promoter studies
title_short	Cre-loxP recombination vectors for promoter studies
title_full	Cre-loxP recombination vectors for promoter studies
title_fullStr	Cre-loxP recombination vectors for promoter studies
title_full_unstemmed	Cre-loxP recombination vectors for promoter studies
title_sort	cre-loxp recombination vectors for promoter studies
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2007
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582007000200013
work_keys_str_mv	AT pedersennina creloxprecombinationvectorsforpromoterstudies AT tuxenpoulsenthomas creloxprecombinationvectorsforpromoterstudies AT skovgaardpoulsenhans creloxprecombinationvectorsforpromoterstudies
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Cre-loxP recombination vectors for promoter studies

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