J. Bacteriol., 04 1996, 2436-2439, Vol 178, No. 8
Copyright © 1996, American Society for Microbiology

Analysis of the spacer DNA between the cyclic AMP receptor protein binding site and the lac promoter

U Flatow, GV Rajendrakumar and S Garges
Developmental Genetics Section, Laboratory of Molecular Biology, National Cancer Institute, Bethesda, Maryland 20892-4255, USA.

The role of the spacer region DNA between the cyclic AMP receptor protein (CRP) site and the RNA polymerase in the lac promoter was examined. We wanted to determine whether the wild-type DNA sequence of this region was an absolute requirement for CRP activation of lac transcription. The sequence of a 9-bp stretch of the spacer, from -41 to -49 relative to the start of transcription, was randomized, and the effect of randomization on lac expression was investigated in vitro and in vivo. We found that the spacer contains no specific sequence determinants for CRP activation of lac transcription; fewer than 1% of the mutants displayed greater than a 50% decrease in CRP activation of lac transcription.

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