Yeast one-hybrid system used to identify the binding proteins for rat glutathione S-transferase P enhancer I.

Ming-xiang Liao; Dong-yuan Liu; Jin Zuo; Fu-de Fang

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Yeast one-hybrid system used to identify the binding proteins for rat glutathione S-transferase P enhancer I.

Author: Ming-Xiang LIAO ¹ ; Dong-Yuan LIU ; Jin ZUO ; Fu-De FANG
Author Information

1. National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100005, China.
Publication Type:Journal Article
MeSH: Animals; Biological Assay; methods; Carrier Proteins; chemistry; isolation & purification; DNA Primers; DNA, Complementary; genetics; Enhancer Elements, Genetic; genetics; Enzyme Induction; Gene Expression Regulation; Gene Library; Glutathione Transferase; genetics; Lung; Rats; Sequence Analysis, DNA; Yeasts
From: Biomedical and Environmental Sciences 2002;15(1):36-40
CountryChina
Language:English
Abstract:
OBJECTIVETo detect the trans-factors specifically binding to the strong enhancer element (GPEI) in the upstream of rat glutathione S-transferase P (GST-P) gene.
METHODSYeast one-hybrid system was used to screen rat lung MATCHMAKER cDNA library to identify potential trans-factors that can interact with core sequence of GPEI(cGPEI). Electrophoresis mobility shift assay (EMSA) was used to analyze the binding of transfactors to cGPEI.
RESULTScDNA fragments coding for the C-terminal part of the transcription factor c-Jun and rat adenine nucleotide translocator (ANT) were isolated. The binding of c-Jun and ANT to GPEI core sequence were confirmed.
CONCLUSIONSRat c-jun transcriptional factor and ANT may interact with cGPEI. They could play an important role in the induced expression of GST-P gene.