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Studied on gene control regions IX. The effect of hypoxanthine-substituted lac operators on the lac operator--lac repressor interaction.
Authors:D G Yansura  D V Goeddel  A Kundu  M H Caruthers
Institution:Department of Chemistry, University of Colorado Boulder, Colo. 80309, U.S.A.
Abstract:Hypoxanthine was substituted for guanine at specific sites in the lac operator DNA by a combination of chemical and enzymatic procedures. The stability of these modified lac operators with wild-type (SQ) and tight binding (QX86) lac repressors was measured. Effects were variable. At some sites insertion of hypoxanthine significantly reduced the stability of the complex whereas at other sites substitution with hypoxanthine did not alter the repressor—operator interaction. In addition, insertion of this analog at two sites increased the stability of the complexes. These changes were used to partially map regions of the lac operator that are in contact with lac represser. The results suggest that lac repressor recognizes the guanine 2-amino group at specific sites in the minor groove of lac operator.
Keywords:RO complex  H  hypoxanthine  dITP  deoxyinosine 5′-triphosphate
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