10 遗传重组和转座子课件.ppt

* a、 RecA 有单、双链 DNA 结合活性,其中，双链结合活性最终导致的也是 RecA 与单链的结合状态 * RecA-mediated strand exchange between partially duplex and entirely duplex DNA generates a joint molecule with the same structure as a recombination intermediate. * The natural migration rate of a holiday junction is relatively slow. In E. coli the proteins Ruv A and Ruv B act as ‘motors’ to drive the migration faster. - ATP dependent process * Λphage进入E.coli后整合到寄主染色体上以及从染色体上切除均 attP 含 POP’三序列240bp（－160～＋80） Sne－－－235bp（－152～＋82） * ? 核心序列O全长15bp，富含A-T无碱基倒转对称性 ? Int结合 ----核心序列的反向位点（在每条链的适当位置上切割） ----结合在att臂上（两位点同时结合使臂位点靠近核心区） * Multiple copies of Int protein may organize attP into an intasome, which initiates site-specific recombination by recognizing attB on free DNA * A composite transposon has a central region carrying markers (such as drug resistance) flanked by IS modules. The modules have short inverted terminal repeats. If the modules themselves are in inverted orientation (as drawn), the short inverted terminal repeats at the ends of the transposon are identical. * 不依赖供体序列与靶位点间序列的同源性 (非同源重组过程 ，不依赖 recA 酶) Regional preference ( 在3kb区域内的随机插入)---------------有的转座元倾向于优先插入某些染色体上的某些大的区段，这些区段可长达3Kb左右 * Figure 15.13 Mu transposition generates a crossover structure, which is converted by replication into a cointegrate. * Nonreplicative transposition results when a crossover structure is released by nicking. This inserts the transposon into the target DNA, flanked by the direct repeats of the target, and the donor is left with a double-strand break * Conservative transposition involves direct movement with no loss of nucleotide bonds; compare with lambda integration and excision. * Transposons of the TnA family have inverted terminal repeats, an internal res site, and three known genes. * Figure 15.4 The direct repeats of target DNA flanking a transposon are generated by the introduction of staggered cuts whose protruding ends are linke