分子生物学课件第六章 DNA的重组与转座.pptVIP

第六章 DNA的重组与转座 2 转座（ transposition ） 2.2 真核生物中的转座成分 真核生物的转座成分根据转座机制目前分为两类： ⑴ 转座机制与细菌的转座子类似 遗传信息： DNA→DNA ? 玉米的Ac-Ds元件、果蝇的P元件和FB元件等 ⑵ 转作机制类似逆转录病毒 遗传信息： RNA→DNA→RNA ? 如：逆转录病毒、果蝇的copia元件、酵母的Ty元件 第六章 DNA的重组与转座 本章结束！！！ * ? 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 * * 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 * 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. * 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 linked to the transposon. 1.2.3 与重组相关的酶 ⑶ RuvA,RuvB和RuvC RuvA四聚体形成的 结构像四个花瓣的花。 Ruv AB复合物结合于 Holliday联结体的模型 第六章 DNA的重组与转座 （左）RuvA四聚体结合到Holliday位点； (RuvA 识别 Holliday 结构的连接点) （中）RuvB六聚体结合到杂合双螺旋的两对面，DNA穿过其 中心， RuvB六聚体的作用像马达，促进超螺旋分叉点通过复合体移动。(具有ATPase活性) （右）RuvC结合到Holli