重组和转座.pptVIP

第一节 重组的分类 一、?同源重组或普遍性重组： Recombination involving reaction between homologous sequences of DNA is called generalized or homologous recombination. 依赖于参与重组的DNA分子在序列上的广泛一致性； 同源性重组可以发生在两条同源DNA上的任何位点； 经常发生于减数分裂期的同源染色体之间。 二、位点特异性重组 Site-specific recombination?(Specialized recombination) 这种重组的特点是重组发生在特异位点，此位点含有短的同源序列,供位点特异性重组酶识别(如 λ噬菌体DNA整合到寄主DNA)； 重组DNA的其它区域不具有同源性 三、转座： refers to the movement of a transposon to a new site in the genome. A transposon?(transposable element) is a DNA sequence able to insert itself (or a copy of itself) at a new location in the genome, without having any sequence relationship with the target locus. 转座分为复制型、非复制及保守型三种类型 四、模板选择（copy choice）性重组 适用于RNA病毒，在这种重组中，聚合酶从一个模板转换到另一个模板来合成RNA，结果新合成的分子将含有两个不同亲本的遗传信息 第二节 同源重组 一、Holliday连接： 在同源染色体的相同位点产生切口 两条染色体的DNA链发生交叉 切口封闭，形成holliday junction (或chi结构) Branch migration（分枝迁移） 在两对同源链的其中任意一对上产生切口，拆分中间体 二、Rec BCD途径 －同源重组的酶学机制 Overview: Rec BCD蛋白同时具有解螺旋酶的活性，使chi位点附近切口的DNA解链 单链区被Rec A蛋白和SSB蛋白覆盖 RecA 蛋白使单链DNA取代双链DNA中的同源部分 D－loop区域的DNA产生切口，新切口的3’端（ the tail of newly nicked DNA ）与另一条DNA的单链区互补配对 DNA连接酶封闭切口，形成Holliday junction RuvA 和RuvB发动迁移反应 RuvC拆分重组中间体 RecBCD: The RecBCD complex has nuclease and helicases activities. It binds to DNA downstream of a chi sequence, unwinds the duplex, and degrades one strand from 3’ –5‘ as it moves to the chi site. The chi site triggers loss of the RecD subunit and nuclease activity. it continues to function as a helicase, now using only the RecB subunit to drive translocation, at about half the previous speed. Rec A: RecA in bacteria has two quite different types of activity: it can stimulate protease activity in the SOS response and can promote base pairing between a single strand of DNA and its complement in a duplex molecule. Both activities are activated by single-stranded DNA in the presence of ATP. The DNA-handling activity of RecA enables a single strand to displace its homologue in a duplex in a reaction that is called single-strand assimilation (单链同