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Published byJob Peters Modified over 9 years ago
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Transcription control elements (DNA sequences) are binding sites for transcription factors, proteins that regulate transcription from an associated promoter. Activators stimulate transcription. Repressors inhibit transcription.
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AATTGGGGCGGGGC CCCCGCCCCGTTAA ( ) n Hybridize and ligate AATTGGGGCGGGGCAATTGGGGCGGGGCAATTGGGGCGGGGC…………………… CCCCGCCCCGTTAACCCCGCCCCGTTAACCCCGCCCCG…………………… AATTGGGGCGGGGCAATTGGGGCGGGGCAATTGGGGCGGGGC…………………… CCCCGCCCCGTTAACCCCGCCCCGTTAACCCCGCCCCG…………………… Specific sequence DNA affinity column Covalently link synthetic, Ligated DNA to a solid support
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Obtain amino acid sequence from isolated transcription factor. Search the cDNA (EST) data base for a cDNA that encodes the transcription factor. Insert this into an expression vector. Assay the ability of the transfected expression vector to stimulate reporter gene expression from a co-transfected reporter plasmid.
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WT or mutant Gal4 Gal4 s WT or mutant Gal4 Expression vector:
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Fig. 11-17
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E. coli LexA yeast Gal4 aa738--823 C N E. coli LexA C N Expression vector for: or LexA binding sites :
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Gal4 reporter LexA reporter LexA vector galactosidase units 400 800 1200 LexA-Gal4(738-823) vector + _ _ _ + + + + _ _ _ _ _ _ + + 1600 2000
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Fig. 11-18
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Fig. 4-17
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Fig. 11-20
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Interface of 434 repressor with operator DNA (lower monomer and dimer Interface)
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Fig. 3-6
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Fig. 2-13
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Sequences of C 2 H 2 zinc-fingers in TFIIIA
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Fig. 11-21 Zinc-finger protein (C 2 H 2 type)
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