Nucleic Acids Research, 1989, Vol. 17, No. 10 3747-3755
© 1989
MOLECULAR BIOLOGY |
Flanking DNA-sequences contribute to the specific binding of cl-repressor and ORI
Department of Biology and McCollum Pratt Institute, The Johns Hopkins University Baltimore, MD 21218, USA
*To whom correspondence should be addressed
Received January 31, 1989. Revised April 12, 1989. Accepted April 12, 1989.
The binding of cl-repressor to a series of mutant operators containing OR1 of the right operator of bacteriophage lambda was investigated. Sites OR2 and/or OR3 were inactivated by either point or deletion mutations. The free energy of binding repressor to OR1 in the wildtype operator, 
G1, is –13.7 ± 0.3 kcal/mol. G1, determined for an OR2– operator created by a single point mutation in OR2 is –13.6 ± 0.2 kcal/mol. In contrast, G1, for the binding of repressor to a cloned synthetic OR1 operator containing only 24 bp of lambda sequence is –12.2 ± 0.1 kcal/mol. When sequence 5' to OR1 is present, the binding affinity increases to –13.0 ± 0.1 kcal/mol. In addition, the proximity of OR1 to a fragment-end decreases G1, from –13.7 to –12.3 ± 0.1 kcal/mol. These results suggest that the DNA sequence outside the 17 bp OR1 binding-site contributes to the specific binding of cl-repressor.
+Present addresses: Department of Biochemistry, The Albert Einstein College of Medicine, Bronx. NY 10461
Present addresses: Department of Molecular Biology and Biochemistry, University of California at Irvine, Irvine, CA 92717, USA
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