MECHANISTIC STUDIES OF GENETIC RECOMBINATION

这项拟议研究的总体目标是为 对基因重组机制的分子理解。这个 方法将涉及物理、生化和 纯化蛋白的酶学性质已知涉及 生物重组过程。这些研究的重点将是 大肠杆菌recA蛋白及其与核酸相互作用的研究 底物，以及与参与的其他蛋白质的相互作用 重组，如SSB蛋白、recBC和DNA解旋酶。整体而言 这些研究的目的是为了理解，在物理机制 水平，这些蛋白质如何相互作用，以及如何与它们的核相互作用 酸性底物以产生重组DNA分子。解决问题的方法 包括结构平衡和结构平衡的物理研究 RecA蛋白与核酸底物结合的动力学研究 RecA蛋白催化DNA等反应的机制 复性和DNA链同化；突变体的生化研究 RecA蛋白变异体，RecA蛋白的蛋白分解片段和 截短的recA基因产物；以及对该基因的物理和酶研究 RecA蛋白与SSB蛋白、recBC、 和DNA解旋酶。

The overall goal of this proposed research is to contribute to the molecular understanding of the mechanism of genetic recombination. The approach will involve the investigation of the physical, biochemical, and enzymatic properties of purified proteins which are known to be involved in the biological recombination process. The focus of these studies will be on the E. coli recA protein, and the interaction with its nucleic acid substrates, as well as the interaction with other proteins involved in recombination, such as SSB protein, recBC, and DNA helicases. The overall objective of these studies is to understand, at the physical mechanistic level, how these proteins interact with each other and with their nucleic acid substrates to produce a recombinant DNA molecule. The approaches to be used include physical investigations of the structural equilibrium and kinetic aspects of binding of recA protein to nucleic acid substrates and of the mechanism of recA protein catalyzed reactions such as DNA renaturation and DNA strand assimilation; biochemical studies of mutant recA protein variants, of proteolytic fragments of recA protein and of truncated recA gene products; and physical and enzymatic studies of the interaction of recA protein with other proteins such as SSB protein, recBC, and DNA helicases.