第九章 基因工程打印稿.pptx

Genetic Engineering 遗传工程 gene cloning 基因克隆 recombination DNA technique 重组DNA技术 Clone（克隆） are identical organisms, cells, or molecules descended from a single ancestor ; 第一节 限制性内切酶与连接酶 Restriction Enzymes and DNA Ligase;Palindrome 回文结构;;Fragments with identical sticky ends can be joined to form recombinant DNA molecule;;;Restriction map（限制酶图谱） Compile the number, order, and distance between restriction sites along a DNA fragment ;2. DNA ligase（DNA连接酶） Catalyzes formation of a covalent bond（共价键） between adjacent 5’-P and 3’-OH termini in a broken double-strand DNA ;;第二节 分子克隆载体Vectors;;pBR322;pUC18;2. Lambda vector（λ载体）;;3. Cosmid （cos 质粒） constructed by inserting the cos sequence of λ, which is necessary for packing phage DNA into phage protein coats, into a plasmid vector;4. Bacterial artificial chromosome ( BAC ) 细菌人工染???体 Derived from F plasmid, can carry inserts of about 300kb;;YAC载体的特点： 两个TEL位点，一个CEN，一个ARS（自主复制序列） 三个酵母菌遗传标记基因 克隆片段大小约1Mb YAc载体缺点：易出现嵌合体，某些克隆不稳定 ;;6. Ti plasmid and plant vectors Agrobacterium tumifaciens 根癌农杆菌 Tumor inducing (Ti) plasmid;第三节 DNA文库 DNA libraries;Calculate the number of clones in a library ;2. cDNA library （cDNA文库）;cDNA libraries represent only the expressed genes in a given cell type, tissue, or stage of embryonic development. ; 第四节 目的DNA的分离 Identifying and Isolating of  Interested DNA;利用核酸探针筛选目的DNA;See movie; Antibody can be used as probes ;;Problems and discussion questions