生物化学本科课件英文版讲授chapter.pptx

Chapter 6  Exploring Genes;Recombinant DNA technology, a new approach to exploring the central molecules of life ;A second foundation is the base-pairing language that mediates nucleic acid recognition. ;some of the benefits of these new methods ;the construction and cloning of novel combinations of genes;the Basic Tools of Gene Exploration基因探索的五大基本工具 1.Restriction-enzyme analysis 限制酶分析 2.Blotting techniques 吸印技术 3.DNA sequencing DNA测序 4.Solid-phase synthesis of nucleic acids 核酸固相合成 5.The polymerase chain reaction (PCR) 聚合酶链反应 A final tool—computer 重组DNA技术的三大基础: 1. 限制酶 2. 碱基配对语言 3. 载体;Restriction Enzymes Split DNA into Specific Fragments;Restriction enzymes, also called restriction endonucleases,限制性核酸酶 recognize specific base sequences in double-helical DNA and cleave both strands of the duplex at specific places;Restriction enzymes are found in a wide variety of prokaryotes. Their biological role is to cleave foreign DNA molecules. ;More than 90 restriction enzymes have been purified and characterized. ;Restriction enzymes are used to cleave DNA molecules into specific fragments that are more readily analyzed and manipulated than the parent molecule;Indeed, complex chromosomes containing hundreds of millions of base pairs can be mapped by using a series of restriction enzymes. ;Restriction Fragments can be Separated by Gel Electrophoresis and Visualized;Small differences between related DNA molecules can be readily detected because their restriction fragments can be separated and displayed by gel electrophoresis;In certain kinds of gels, fragments differing in length by just one nucleotide out of several hundred can be distinguished. Moreover, entire chromosomes containing millions of nucleotides can now be separated on agarose gels by applying pulsed electric fields脉冲电场 in different directions. (大片段DNA可因所施加的电流的角度连续改变而得以分离) Bands or spots of radioactive DNA in gels can be visualized by autoradiography放射自显影. Alternatively, a gel can be stained with ethidium