Schaper S, Messer W
Max-Planck-Institut für Molekulare Genetik, Berlin-Dahlem, Germany.
J Biol Chem. 1995 Jul 21;270(29):17622-6. doi: 10.1074/jbc.270.29.17622.
Equilibrium and kinetic rate constants were determined for the binding of the initiator protein DnaA of Escherichia coli to its binding site, the non-palindromic 9-bp DnaA box, using gel retardation techniques. The dissociation constant for specific binding was between 1 and 50 nM for individual DnaA boxes on 21-bp double-stranded oligonucleotides. Only DnaA boxes of the sequence TT(A/T)TNCACA resulted in specific fragment retention. Both the 9-bp consensus sequence and flanking sequences determined the binding efficiency. One DnaA monomer was found to bind to a DnaA box and to induce a bend of about 40 degrees.
运用凝胶阻滞技术，测定了大肠杆菌起始蛋白DnaA与其结合位点（非回文9碱基对DnaA框）结合的平衡常数和动力学速率常数。对于21碱基对双链寡核苷酸上的单个DnaA框，特异性结合的解离常数在1至50 nM之间。只有序列为TT(A/T)TNCACA的DnaA框能导致特异性片段保留。9碱基对共有序列和侧翼序列均决定了结合效率。发现一个DnaA单体可与一个DnaA框结合，并诱导约40度的弯曲。
