Accurate and quantitative transcription of the tk gene requires sequence elements that reside within 110 nucleotides of 5'-flanking DNA. We have determined the boundaries of a hypersensitive region in 5'DNA flanking the tk gene by analyzing the relative sensitivity of specific restriction sites clustered in this region. Five different restriction enzymes, recognizing over 50 sites in the promoter region of the tk gene, each show a preferential cleavage in nuclei at a restricted number of sites residing between positions -4 and -182. Thus the tk promoter sequences are contained entirely within a hypersensitive region. Analysis of this site in tk+ transformants, a tk- mutant and tk+ rerevertant indicates that expression of tk RNA is correlated with structural alterations in the tk promoter.
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