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A 16 base-pair palindrome upstream of the Agrobacterium tumefaciens octopine synthase (ocs) gene functions as a positive regulatory element in plant cells (Ellis et al. (1987) EMBO J. 6, 3203-3208; Fromm et al. (1989) Plant Cell 1, 977-984). We have converted it into a negative element by locating two copies flanking the TATA-box of the constitutively expressed CaMV 35S promoter. The reduced promoter activity is very likely due to sterical hindrance of the ocs binding protein (OCSTF) x ocs complex with the transcription initiation complex. We propose that this type of constructs can be used for the identification of recognition sites for DNA-binding proteins which are labile in vitro as well as for determining the DNA-binding activity of a trans-acting factor in vivo.
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