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Extra info for Current Topics in Microbiology and Immunology / Ergebnisse der Mikrobiologie und Immunitatsforschung
1974) found in addition that certain promoter mutations in P L or in P R , which are known to decrease the affinity of polymerase for these sites, also abolish the Hind II cleavage site in the corresponding operator. Revertants which have recovered promoter activity also recover the restriction site. Clearly the two sites must have elements in common. WALZ and PIRROTTA (1975) have isolated and sequenced the DNA fragment originating from the OR region, which the RNA polymerase can protect against nuclease digestion.
PIRROTTA: 34 Table 1. Constants regulating repressor-operator interaction. "[1/2 is dissociation half-life of repressor-operator complex. , 1971, and unpublised data of P. CHADWICK) kforward 7 X 10-9 mole liter- 1 2 X 10 22 mole 2 liter- 2 3 X 10-14 mole liter-' 3 X 10'0 mole-1 liter sec-1 175 min 7 min same conclusion. In both cases the analysis shows a dependence on the square of repressor concentration. These conclusions, however, could equally well apply to a dimer-tetramer equilibrium. B. The Binding Reaction The filter binding technique has been successfully employed by RIGGS and BOURGEOIS (1968) to study the parameters of the lac repressor-operator interaction.
Wu et aI. 2 M KCI suffices to dissociate 95 % of the complexes. Increasing temperature and pH also weaken the binding. 5. The temperature effect occurs at the level of interaction with the operator rather than on the monomer-oligomer equilibrium, since it is due to an increase in the rate of dissociation of the repressor-operator complex. 05 M KCI the half-life of the complex is 175 minutes, while at 20° C it becomes The A Repressor and its Action 35 7 minutes, decreasing by a factor of 5 for every 10° C in crease in temperature.