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遗传重组的起始:由recA蛋白促进的双链DNA分子之间的同源配对。

Initiation of genetic recombination: homologous pairing between duplex DNA molecules promoted by recA protein.

作者信息

Cassuto E, West S C, Mursalim J, Conlon S, Howard-Flanders P

出版信息

Proc Natl Acad Sci U S A. 1980 Jul;77(7):3962-6. doi: 10.1073/pnas.77.7.3962.

Abstract

recA protein has been shown to promote hydrogen bonding between single-stranded DNA fragments and duplex DNA molecules homologous to them. However, genetic and biochemical evidence indicates that genetic exchanges generally take place between duplex molecules. We therefore chose to study the interactions promoted by recA protein between intact duplex DNA molecules and molecules containing gaps that are believed to increase the frequency of genetic exchanges. In the present paper, we show that incubation of intact and gap-containing plasmid DNA in the presence of recA protein leads to homologous pairing between duplex molecules which can be detected by centrifugation analysis and electron microscopy. The reaction is completely dependent on an active recA gene product, on genetic homology between the DNA species involved, and on the presence of ATP; under certain conditions, its efficiency can be increased considerably by the presence of the single-stranded DNA binding protein of Escherichia coli.

摘要

RecA蛋白已被证明可促进单链DNA片段与与其同源的双链DNA分子之间形成氢键。然而,遗传学和生物化学证据表明,基因交换通常发生在双链分子之间。因此,我们选择研究RecA蛋白促进完整双链DNA分子与含有缺口的分子之间的相互作用,据信这些缺口会增加基因交换的频率。在本文中,我们表明,在RecA蛋白存在下孵育完整的和含缺口的质粒DNA会导致双链分子之间的同源配对,这可以通过离心分析和电子显微镜检测到。该反应完全依赖于活性RecA基因产物、所涉及的DNA种类之间的遗传同源性以及ATP的存在;在某些条件下,大肠杆菌单链DNA结合蛋白的存在可显著提高其效率。

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