分子生物学课件chapter5dna的变性与修复.pptxVIP

Chapter 5 the mutabiligyt and repair of DNA Chapter 9 of the book;Where does the mutation come from? DNA replication Chemical damage to the gene material Insertions caused by DNA elements; 突变体(mutant)：携带突变的生物个体或群体或株系; ● Chromosome mutation ;Point mutation－－－碱基替代、碱基插入、碱基缺失;Replication errors and their repair;DNA mismatch;DNA polymerase Helicase SSB 外切核酸酶 (Ⅰ和Ⅶ) ligase;Mismatch repair pathway for the repair of replication errors;★ the transient hemimethylation in the DNA replication;a、DNA/MutS complex recruits MutL;外切核酸酶Ⅰ切割切点的5’端（错配碱基在切点的5’端） ----------------Ⅶ----------------3’端（--------------------------3’端）;How to repair DNA damage?;1. Alkylation (烷化剂) 亚硝酸（nitrous acid HNO2） 羟氨（hydroxylamine HA） 甲磺酸乙酯（ethyl methanesulfonate EMS） N-甲基-N’-硝基-N-亚硝基胍 （N-methyl-N’-nitro-N-nitrosoguanidion NNG）;NH2OH (Hydroxylamine HA 羟胺);2. Radiation Ultraviolet light, γ radiation , x-ray ;;3、Base analog;5-溴尿嘧啶 5-Bromine Uracil ;;吖啶橙 (Acridine Orange AO) ;The systems repairing damage to DNA Excision repair systems Recombinantional repair;1. Direct repair of DNA;photoreactivation;2. Methyl group removal;甲基转移酶;3.核苷酸切除修复;;;● 与Rec-A蛋白引起的重组(strand transfer)有关; 重组修复 (链转移修复);RecA蛋白首先与单链DNA结合（需消耗ATP）。于是RecA蛋白即被活化而可将双螺旋解旋和分离，同时企图将它结合的单链与被解旋区域退火，如此继续下去，直到找到互补顺序。只要一旦有一小部份被真正“退火”，ATP供应的能量就会继续驱使配对反应趋于完成。当新的杂交双链形成时，RecA蛋白即从原来的单链掉下来。;Repair double-strand break;5. SOS respones (SOS 修复) The SOS response is a global response to DNA damage in which the cell cycle is arrested and DNA repair and mutagenesis are induced. The SOS uses the RecA protein (Rad51 in eukaryotes). The RecA protein, stimulated by single-stranded DNA, is involved in the inactivation of the LexA repressor thereby inducing the response. It is an error-prone repair system. ;Function of RecA protein ;mechanism of SOS response; SOS 修复只是SOS反应的一部分;当DNA复制受阻/ DNA damaged;当DNA复制度过难关后;第三节 限制和修饰; 1、 限制－修饰现象(restriction and modificaion);结论：; 自己DNA: 限制模式相同的DNA