Khoury A M, Lee H J, Lillis M, Lu P
Department of Chemistry, University of Pennsylvania, Philadelphia 19104.
Biochim Biophys Acta. 1990 Sep 10;1087(1):55-60. doi: 10.1016/0167-4781(90)90120-q.
The interaction of the E. coli lac operon repressor with its operator DNA has been directly examined as a function of the length of operator-containing DNA. The apparent bimolecular association rate constants were calculated as ka = (kd/KD), where the dissociation equilibrium constant, KD and the dissociation rate constant, kd, were measured by nitrocellulose filter adsorption assays. The values obtained for the overall association rate constants are compared with theoretical association rate curves for specific mechanisms. Association of the repressor with short operator containing DNA fragments (less than 70 base pairs) occurs at rates expected of three-dimensional diffusion. Our data also imply that at longer DNA lengths a combination of three-dimensional diffusion with one-dimensional sliding along with hopping and/or intersegment transfer must be involved to facilitate the repressor operator association.
已经直接研究了大肠杆菌乳糖操纵子阻遏物与其操纵基因DNA的相互作用,该相互作用是含操纵基因DNA长度的函数。表观双分子缔合速率常数计算为ka = (kd/KD),其中解离平衡常数KD和解离速率常数kd通过硝酸纤维素滤膜吸附测定法测量。将获得的总缔合速率常数的值与特定机制的理论缔合速率曲线进行比较。阻遏物与含短操纵基因的DNA片段(小于70个碱基对)的缔合以三维扩散预期的速率发生。我们的数据还表明,在更长的DNA长度下,必须涉及三维扩散与一维滑动以及跳跃和/或片段间转移的组合,以促进阻遏物与操纵基因的缔合。