Cassuto E, West S C, Howard-Flanders P
EMBO J. 1982;1(7):821-5. doi: 10.1002/j.1460-2075.1982.tb01254.x.
RecA protein has been shown to promote the formation of joint molecules between intact duplex DNA and homologous gapped DNA. When examined by electron microscopy, such joint molecules display a junction that is, in most cases, distant from the site of the gap. This led us to test whether the observed location of the joint was due to pairing at the gap followed by branch migration, or whether recA-promoted pairing could also take place between duplex homologous regions away from the gap. To test the latter possibility, intact duplex DNA was incubated with DNA which contained a gap in a region of non-homology. Joint molecules were detected by filter binding assay and by electron microscopy at about one-third of the yield observed for fully homologous molecules. These results indicate that initial homologous pairing promoted by recA protein is not restricted to the single-stranded region in the gap but can also take place in regions where both molecules are duplex.
RecA蛋白已被证明能促进完整双链DNA与同源缺口DNA之间形成联合分子。通过电子显微镜检查时,此类联合分子显示出一个连接点,在大多数情况下,该连接点远离缺口位点。这促使我们去测试观察到的联合位置是由于在缺口处配对后发生分支迁移,还是RecA促进的配对也能在远离缺口的双链同源区域之间发生。为了测试后一种可能性,将完整的双链DNA与在非同源区域含有一个缺口的DNA一起孵育。通过滤膜结合试验和电子显微镜检测到联合分子,其产量约为完全同源分子观察产量的三分之一。这些结果表明,RecA蛋白促进的初始同源配对不仅限于缺口中的单链区域,也能在两个分子均为双链的区域发生。