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Chinese Journal of Biotechnology ; (12): 343-346, 2007.
Article Dans Chinois | WPRIM | ID: wpr-328026

Résumé

To establish a new high-throughput screening model for the agonist of PPARdelta, PPARdelta gene was obtained by reverse transcriptase-polymerase chain reaction (RT-PCR), and subcloned to pGEM-T Vector for sequencing, then the PPARdelta fragment was excised by restriction enzymes, and inserted into pTARGET Vector to construct expression vector pTARGET-ppARdelta. Insert three copies of PPRE into pGl3-promoter vector to construct expression vector pGl3-PPRE x 3-luc. The vector pTARGET-ppARdelta was transiently cotransfected with pGl3-PPRE x 3-luc into different cell lines to assay the expression levels of luciferase. The PPARdelta agonist screening model was established and optimized. Bezafibrate and linoleic acid can induce the expression of luciferase significantly and in a dose-dependent manner. This method can be used for high throughput screening for the agonist of PPARdelta, which might become lead compounds for new anti-atheroscleriosis or anti-adiposity drugs.


Sujets)
Animaux , Humains , Souris , Cellules 3T3 , Bézafibrate , Pharmacologie , Lignée cellulaire , Relation dose-effet des médicaments , Évaluation préclinique de médicament , Méthodes , Vecteurs génétiques , Chimie , Génétique , Cellules HeLa , Acide linoléique , Pharmacologie , Lipides , Chimie , Luciferases , Génétique , Métabolisme , Récepteur PPAR delta , Génétique , Protéines de fusion recombinantes , Génétique , Métabolisme , Transfection , Méthodes
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