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DOROTHY BUCK |
The Topological Aspects of Rearrangement Reactions Involving DNA |
This series of experiments utilizes topological methods to determine how
enzymes involved in DNA recombination affect the architecture of DNA. For
DNA longer than 200bp, we determine if the recombinase Flp introduces writhe,
by considering the product when run through a two-dimensional electrophoretic
gel. By utilizing multiple sites, we yield additive writhe, amplifying
our results. Next, given Gin or resolvase, with known topological effects,
we build a synaptic complex consisting of Flp and Gin (or resolvase). By
comparing their product to know knots and links, we can determine the sign
of the strand rotation of the recombination enzyme. Having established
the sign, we can then probe the architecture of the synaptic complex of
unknown enzymes. by adding the rotation site to more complexes, such as
that of Mu transposase, we can deduce the synapse topology of other recombinases. |
References: |
Cozzarelli, N.R. (1992). Proceedings of Symposia in Applied Mathematics.
45: 1-16. Schwartz, C.J.E. and Sadowski, P.D. (1990). J.Mol.Biol. 216: 289. Chen, J.W., Evans, B.R., Rosenfeldt, H. and Jayaram, M. (1992). Gene. 119: 37. Chen, J.W., Lee, J. and Jayaram, J. (1992). Cell. 69:547. Sumners, D.W., Ernst, C., Spengler, S.J. and Cozzarelli, N.R. (1995). Quarterly Reviews of Biophysics. 28, #3: 255.
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