DISCOVERY OF A PREDICTED DNA KNOT SUBSTANTIATES A MODEL FOR SITE-SPECIFIC RECOMBINATION
DISCOVERY OF A PREDICTED DNA KNOT SUBSTANTIATES A MODEL FOR SITE-SPECIFIC RECOMBINATION
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DOI：
10.1126/science.2990045
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发表时间：
1985-01-01
期刊：
影响因子：
56.9
通讯作者：
COZZARELLI, NR
中科院分区：
文献类型：
--
作者：
WASSERMAN, SA;DUNGAN, JM;COZZARELLI, NR
通过拓扑学方法研究了 Tn3 解离酶介导的位点特异性基因重组机制。突触和链交换的详细模型的外推可以预测具有特定打结结构的额外 DNA 产物的形成。体外反应的 DNA 的二维凝胶电泳显示出产物，.apprx。总量的0.1%，具有适当的流动性。暗示了一种通过 EM 确定 DNA 拓扑结构的技术，所需的 DNA 少于一纳克。结的结构符合预测，为模型提供了有力的证据，显示了拓扑方法的强大功能。
The mechanism of site-specific genetic recombination mediated by Tn3 resolvase was investigated by a topological approach. Extrapolation of a detailed model of synapsis and strand exchange predicts the formation of an additional DNA product with a specific knotted structure. Two-dimensional gel electrophoresis of DNA reacted in vitro revealed a product, .apprx. 0.1% of the total, with the appropriate mobility. A technique for determining DNA topology by EM was implied such that less than a nanogram of DNA was required. The structure of the knot was as predicted, providing strong evidence for the model and showing the power of the topological method.