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.
AATTGGGGCGGGGC CCCCGCCCCGTTAA( )n
Hybridize andligate
AATTGGGGCGGGGCAATTGGGGCGGGGCAATTGGGGCGGGGC…………………… CCCCGCCCCGTTAACCCCGCCCCGTTAACCCCGCCCCG……………………
AATTGGGGCGGGGCAATTGGGGCGGGGCAATTGGGGCGGGGC…………………… CCCCGCCCCGTTAACCCCGCCCCGTTAACCCCGCCCCG……………………
Specific sequence DNA affinity column
Covalently link synthetic,Ligated DNA to a solid support
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.
Gal4 reporterLexA reporter
LexA vector
g
alac
tosi
das
e u
nit
s
400
800
1200
LexA-Gal4(738-823) vector
+___
+ +
++
_
___
_
_+
+
1600
2000
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