A phage display combined with DNA affinity magnetic system can be applied to a screening of DNA binding proteins, such as transcription factors / Un fago pantalla combinado con un sistema de afinidad magnético al ADN se puede aplicar al exámen de las proteínas de unión al ADN, tales como los factores de transcripción
Electron. j. biotechnol
;
13(1): 14-15, Jan. 2010. ilus, tab
Article
Dans Anglais
| LILACS
| ID: lil-559597
ABSTRACT
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.
Texte intégral:
Disponible
Indice:
LILAS (Amériques)
Sujet Principal:
Facteurs de transcription
/
Arabidopsis
Type d'étude:
Etude diagnostique
/
Étude de dépistage
Limites du sujet:
Animaux
langue:
Anglais
Texte intégral:
Electron. j. biotechnol
Thème du journal:
Biotechnologie
Année:
2010
Type:
Article
/
descriptif de projet
Pays d'affiliation:
Japon
Institution/Pays d'affiliation:
Okayama University/JP
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