A phage display combined with DNA affinity magnetic system can be applied to a screening of DNA binding proteins, such as transcription factors

Here we introduce a new approach for the screening of DNA binding proteins, using a phage library based on a phage display technique. In principal, a complementary DNA (cDNA) library based on the recombinant bacteriophage T7 expressing target proteins on its capsid (phage display) is constructed. Th...

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Autores principales:	Yulita,Kusumadewi Sri, Kouno,Takafumi, Ezaki,Bunichi
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2010
Materias:	AtGST11 gene biopanning DNA binding proteins T7 phage differential display transcription factors
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000100014
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spelling	oai:scielo:S0717-345820100001000142010-04-28A phage display combined with DNA affinity magnetic system can be applied to a screening of DNA binding proteins, such as transcription factorsYulita,Kusumadewi SriKouno,TakafumiEzaki,Bunichi AtGST11 gene biopanning DNA binding proteins T7 phage differential display transcription factors Here we introduce a new approach for the screening of DNA binding proteins, using a phage library based on a phage display technique. In principal, a complementary DNA (cDNA) library based on the recombinant bacteriophage T7 expressing target proteins on its capsid (phage display) is constructed. These phage particles are hybridized with a biotinylated target DNA fragment which is immobilized on the surface of streptavidin paramagnetic particle (SA-PMP). The phage particles are released from the target DNA fragment by a nuclease treatment and the recovered phages are used to the next round of hybridization. These processes are repeated three times to amplify the target phages in the population. This simple method is faster, and more systemic than other current methods (e.g. yeast one hybrid system). As a proof of this principle, we tried to isolate transcription factors which specifically bind to the promoter region of the Arabidopsis thaliana AtGST11 gene. Two obtained candidates, RING zinc finger protein and AtHB6, showed DNA binding activity to the AtGST11 promoter region. We could validate that our new application of phage display is a superior method for isolation of DNA binding proteins with a broad range of potential applications.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.13 n.1 20102010-01-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000100014en10.4067/S0717-34582010000100014
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	AtGST11 gene biopanning DNA binding proteins T7 phage differential display transcription factors
spellingShingle	AtGST11 gene biopanning DNA binding proteins T7 phage differential display transcription factors Yulita,Kusumadewi Sri Kouno,Takafumi Ezaki,Bunichi A phage display combined with DNA affinity magnetic system can be applied to a screening of DNA binding proteins, such as transcription factors
description	Here we introduce a new approach for the screening of DNA binding proteins, using a phage library based on a phage display technique. In principal, a complementary DNA (cDNA) library based on the recombinant bacteriophage T7 expressing target proteins on its capsid (phage display) is constructed. These phage particles are hybridized with a biotinylated target DNA fragment which is immobilized on the surface of streptavidin paramagnetic particle (SA-PMP). The phage particles are released from the target DNA fragment by a nuclease treatment and the recovered phages are used to the next round of hybridization. These processes are repeated three times to amplify the target phages in the population. This simple method is faster, and more systemic than other current methods (e.g. yeast one hybrid system). As a proof of this principle, we tried to isolate transcription factors which specifically bind to the promoter region of the Arabidopsis thaliana AtGST11 gene. Two obtained candidates, RING zinc finger protein and AtHB6, showed DNA binding activity to the AtGST11 promoter region. We could validate that our new application of phage display is a superior method for isolation of DNA binding proteins with a broad range of potential applications.
author	Yulita,Kusumadewi Sri Kouno,Takafumi Ezaki,Bunichi
author_facet	Yulita,Kusumadewi Sri Kouno,Takafumi Ezaki,Bunichi
author_sort	Yulita,Kusumadewi Sri
title	A phage display combined with DNA affinity magnetic system can be applied to a screening of DNA binding proteins, such as transcription factors
title_short	A phage display combined with DNA affinity magnetic system can be applied to a screening of DNA binding proteins, such as transcription factors
title_full	A phage display combined with DNA affinity magnetic system can be applied to a screening of DNA binding proteins, such as transcription factors
title_fullStr	A phage display combined with DNA affinity magnetic system can be applied to a screening of DNA binding proteins, such as transcription factors
title_full_unstemmed	A phage display combined with DNA affinity magnetic system can be applied to a screening of DNA binding proteins, such as transcription factors
title_sort	phage display combined with dna affinity magnetic system can be applied to a screening of dna binding proteins, such as transcription factors
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2010
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582010000100014
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A phage display combined with DNA affinity magnetic system can be applied to a screening of DNA binding proteins, such as transcription factors

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