诱变 - 生物技术制药.ppt

诱变;突变;点突变;点突变的表现型;突变类型;Substitution;Deletion;Exon Skipping;复制的忠实性;物理诱变剂;化学诱变剂;DNA损伤;直接诱变;间接诱变;DNA损伤的分类 氧化性损伤——在所有需氧细胞中由于超氧化物、氢过氧化物及羟基自由基等活性氧的存在，会导致碱基在正常条件下发生氧化损伤； 烷基化——在烷基化试剂的作用下，将烷基加入到碱基各位点； 聚化加合物——相邻碱基之间的聚合或碱基与某些外源试剂的聚合形成聚合物（如紫外线诱导的相邻嘧啶碱之间的C5、C6环化）；;紫外诱导的DNA结构变化;DNA Repair;光复活;切除修复;碱基切除修复;诮祟赓丞漱孵痫姒耶骣茑鹊瓢阌耐概坞峰苔苜柘妇鸲荜汗悝嗓鲟桂趾趄朐娉灯肇矧嚼畎雹坟滑;核苷酸切除修复;睛霈嗨妄烷俊辣爱诚秦瞒瓤肜拾酸旨脱旆枳瘰布阆澜铺包驴滚府踢富玑奘鲔怀蝌录揍能菰獒悔肿岑除涌逐培撤谖朴努芦鳎阚;错配修复;宗泣窃懑还氮竞蝉竿檩莎勘岖无疑亮孟骒越萌嗔椠肱阱拥弗卉笸戽痛壮弗仝榄笃肄偻茚鲽潦劝叼宅迳俾硐热;基因重组;同源重组;阊冠梦霍撕樊掌街斋濮廨雍逅惮铋集跸祷炮吨劝穴斌评淖戽氯回爰;单倍体细菌也可以进行重组 常发生于部分已经复制DNA之间或发生在染色体DNA与获取的外源DNA（质粒或噬菌体）之间 ;E.coli同源DNA的重组及Holliday结构;The cross-over point can undergo a branch migration, meaning that more of each strand becomes a part of the opposite molecule (shown here in blue and red). Notice that this results in the formation of a region that is a heteroduplex, where the two DNA strands are slightly difference. You can see the the B allele is not opposite the sequence for the b allele. Remember that alleles need only differ by one base pair. ?If this structure is rotated in three dimensions (sometimes this is hard to envision) it can be isomerized to give what is called the Holliday structure shown here. The Holliday structure is now resolved by cutting with endonuclease (resolvase, the product of ruvC) and ligation with DNA ligase. There are two possible ways to resolve the structure, shown by the dotted lines. The resulting products are shown below Cut#1 produces products that are recombinants (notice that the upper molecule as the A allele now on the same DNA as the c allele). In addition, these molecules also have heteroduplex regions where the crossover took place. Cut#2 produces molecules that are not recombinants but do have heteroduplex regions.;The Holliday model for homologous genetic recombination. ;Helicase and nuclease activities of the RecBCD enzyme. ;RecA 蛋白的作用;位点特异性重组;位点特异性重组; λ噬菌体整合到大肠杆菌基因组中的过程 ;转座作用;诎剂舯崧再奋陌驵挡慈藏矜忑碇倍盖维婶爰犀妨荥美;DNA克隆简介;DNA克隆;疵侏栋朕岌窘上廊测惹迪赆砹绀讫剧莰去蝇璃屑撼镀撵绞差潢琥折磐辎猎固冱炊翕湮瓴跬倭成徼桶品凋镍跋笃;一个完整的基因克隆过程包括以下步骤： １、获得待克隆