Biology – Quantitative Biology – Quantitative Methods
Scientific paper
2008-11-24
Biology
Quantitative Biology
Quantitative Methods
34 pages, 10 figures, accepted by Biophysical Journal
Scientific paper
We have investigated the binding interaction between the bacteriophage lambda repressor CI and its target DNA using total internal reflection fluorescence microscopy. Large, step-wise changes in the intensity of the red fluorescent protein fused to CI were observed as it associated and dissociated from individually labeled single molecule DNA targets. The stochastic association and dissociation were characterized by Poisson statistics. Dark and bright intervals were measured for thousands of individual events. The exponential distribution of the intervals allowed direct determination of the association and dissociation rate constants, ka and kd respectively. We resolved in detail how ka and kd varied as a function of 3 control parameters, the DNA length L, the CI dimer concentration, and the binding affinity. Our results show that although interaction with non-operator DNA sequences are observable, CI binding to the operator site is not dependent on the length of flanking non-operator DNA.
Cox Edward C.
Golding Ido
Guo Ling
Ong N. P.
Wang Yufang
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