第三章讲 基因操作.ppt

第三章: 基因操作;DNA克隆 ;The first cloni;DNA克隆的基本步骤(I);DNA克隆的基本步骤(II);宿主与载体Hosts ve;限制性内切酶(Restrict;识别位点(Recognitio;无标题;限制性内切酶和DNA连接酶在D;碱性磷酸酶(Alkaline ;自主复制 (autonomou;多克隆位点Mutiple cl;载体的类型 克隆载体(Clon;克隆载体(Cloning ve;克隆与筛选CaCl2Elect;??互补 ??compleme;λ Phage life cy;coscos 20-;噬菌斑(Plaques);5‘-CGGGGCGGCGAC;无标题;粘粒(cosmid)载体克隆;YAC载体克隆telomere;*Selection in S;M13 phage- A fi;M13 phage vecto;无标题;Hybrid plasmid-;无标题;依据DNA片段的大小选择合适的;除了克隆载体的特性外,还具有启;ori;pET vectors 克隆基;1. lacUV-5 ;His-taq and Ni-;整合载体(Integratio;- Vectors for t;农杆菌与Ti质粒;Baculovirus vec;无标题;Mammalian viral;穿梭载体(Shuttle ve;亚克隆(Subcloning);Preparation of ;无标题;质粒DNA转化 (plasmi;转染 (Transfectio;DNA文库(DNA libra;基因组文库prepared f;Purify genomic ;cDNA文库构建 mRN;反转录与cDNA合成;文库的大小- to ensur;for a probabili;文库筛选 标记探针: Radi;How to design a;biological acti;琼脂糖凝胶电泳(Agarose;1 2 3 ;无标题;DNA片段大小的估计;DNA片段的纯化与回收inse;核酸杂交示意图;Southern blotti;转膜装置与放射自显影;Northern blotti;Hybrid arrest a;Southern and No;Labeling of DNA;End labeling5’-;DNA片段标记---Nick ;Nonradioative T;Colony and plaq;无标题;无标题;Western BlotPri;ApplicationsBlo;聚合酶链式反应(polymer;The PCR cycleDe;Primers1. PCR ;兼并引物Degenerate ;Reverse transcr;质粒DNA的制备 Growth;无标题;mRNA isolation-;1. Traditionall;染色体游走技术Chromoso;染色体游走技术原理;染色体游走技术的应用;逆向PCRInverse PC;单酶切DNA片段插入的方向问题;How to identify;无标题;无标题;Analysis of a c;限制性图谱分析Restrict;Combinational e;部分消化(Partial di;限制性图谱分析实例;无标题;DNA sequencingM;G DMS;DNA Sequencing ;dNTPddNTPLacks ;无标题;Sanger法的基本步骤;Automatic seque;RNA sequencingI;RNA specific cl;Sequence databa;Analysis of seq;Genome sequenci;* S1 nuclease ;3‘-end filling;引物延伸Primer Exte;凝胶阻滞分析(Gel reta;DNase I 印迹分析(DN;Mutagenesis of ;Site-directed m;Mutagenesis by ;base substituti;克隆技术的应用 DNA指纹分析;Recombinant pro;Genetically mod;无标题;???杆菌基因介导植物育种;firefly lucifer;1996.7.5-2003.2;DNA指纹技术(DNA fin;无标题;无标题;实验步骤;DNA指纹技术的应用I. ;D1: Mom and Da;强奸案罪犯鉴定;Medical diagnos;Gene therapyAtt;作业题重组DNA分子构建中,常