基因工程第一章 (2)精.ppt

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1、基因工程第一章基因工程第一章基因工程第一章基因工程第一章第1页，本讲稿共34页vXudong Su 苏旭东苏旭东vOffice:D-1320,Biological engineering vTel:7528192,13930891449vEmail:vMaterials can be obtained from:-http:/ -http:/ -http:/ -http:/ 第2页，本讲稿共34页Reference books:v吴乃虎，基因工程原理吴乃虎，基因工程原理 v孙乃恩孙乃恩，分子遗传学，分子遗传学v楼士林楼士林，基因工程，基因工程 vT.A.Brown,Gene Cloning a

2、nd DNA Analysis第3页，本讲稿共34页Chapter one An introduction of genetic engineering第4页，本讲稿共34页Chapter one Hereditary（遗传）（遗传）factor1.一、一、naissance（诞生）（诞生）of genetic engineering 1900 Mendel assumed that each heritable property(or character or trait)of an organism might be controlled by a factor-genetic mater

3、ial based on his famous out-cross(异型杂交异型杂交)experiment.Mendel第5页，本讲稿共34页Nomenclature of Gene2.Chapter one 1909 Wilhelm Johannsen,a Danish botanist,plant physiologist and geneticist.Elemente der exakten Erblichkeitslehre(The Elements of Heredity).It was in this book introduced the term gene.1910 Ameri

4、can scientist Morgan suggested that gene was mapped on chromosomes.JohannsenMorgan第6页，本讲稿共34页Genes reside on DNA molecule3.Chapter one In 1944,the American geneticist Oswald T.Avery and his colleagues clearly demonstrated that DNA carried the genetic information in bacteria,genetic material is DNA,n

5、ot protein.Averys work was not generally accepted until 1952,when American scientists Alfred Hershey and Martha Chase showed that the hereditary material of the T2 virus,a virus that infects bacteria,is also DNA.Avery第7页，本讲稿共34页Discovery of the helix DNA structure4.1952 Watson and Crick “This struct

6、ure has novel features which are of considerable biological interest.It appeared in April 1953 in the scientific paper where James Watson and Francis Crick presented the structure of the DNA-helix,the molecule that carries genetic information from one generation to the other.Chapter one 第8页，本讲稿共34页R

7、ecombinant DNA technology and gene cloning5.In 1970s，Berg used a restriction enzyme to isolate a gene from a human cancer-causing monkey virus.Then,he used ligase to join the section of virus DNA with a molecule of DNA from the bacterial virus lambda,creating the first recombinant DNA molecule.Chapt

8、er one Berg第9页，本讲稿共34页 Berg realized the dangers of his experiment and temporarily terminated it before the recombinant DNA molecule was added to E.Coli,where it would have been quickly reproduced.Following the termination of his experiment,he proposed a one year moratorium(延延期）期）on recombinant DNA

9、studies.Berg later resumed his studies into recombinant DNA techniques,and was awarded the 1980 Nobel Prize in chemistry.His discoveries laid the foundation for field of genetic engineering,and the modern biotechnology industry.Chapter one 第10页，本讲稿共34页 In 1973,Cohen and Boyer created the first recom

10、binant DNA organism using recombinant DNA techniques pioneered a year earlier by Paul Berg.Recombinant DNA,also called gene splicing(粘接）粘接）.Cohens implementation of the technique laid the foundations for todays modern genetic engineering industry.genetic engineering was born6.Chapter one Cohen第11页，本

11、讲稿共34页二、二、conceptions of genetic engineering1.genetic engineering:(classical)基因工程是指将一种或多种生物体（供体）的基因与载体在体外基因工程是指将一种或多种生物体（供体）的基因与载体在体外进行拼接重组，然后转入另一种生物体（受体内），使外源基因在受进行拼接重组，然后转入另一种生物体（受体内），使外源基因在受体细胞中进行复制和表达，定向地创造出生物的新性状。体细胞中进行复制和表达，定向地创造出生物的新性状。Chapter one vv第一代基因工程第一代基因工程第一代基因工程第一代基因工程 蛋白多肽基因的高效表达蛋白多

12、肽基因的高效表达蛋白多肽基因的高效表达蛋白多肽基因的高效表达 经典基因工程经典基因工程经典基因工程经典基因工程vv第二代基因工程第二代基因工程第二代基因工程第二代基因工程 蛋白编码基因的定向诱变蛋白编码基因的定向诱变蛋白编码基因的定向诱变蛋白编码基因的定向诱变 蛋白质工程蛋白质工程蛋白质工程蛋白质工程vv第三代基因工程第三代基因工程第三代基因工程第三代基因工程 代谢信息途径的修饰重构代谢信息途径的修饰重构代谢信息途径的修饰重构代谢信息途径的修饰重构 途径工程途径工程途径工程途径工程 vv第四代基因工程第四代基因工程第四代基因工程第四代基因工程 基因组或染色体的转移基因组或染色体的转移基因组或染

13、色体的转移基因组或染色体的转移 基因组工程基因组工程基因组工程基因组工程第12页，本讲稿共34页Genetic engineering micro plant animal质粒质粒plasmid基基因因重组重组DNArecombinant DNAChapter one 第13页，本讲稿共34页 DNA RNA ProteinReverse transcription Transcription Translation RNA replicationRNA editing Replicationcentral dogma(中心法测中心法测):Chapter one 第14页，本讲稿共34页2.C

14、ontent of genetic engineeringvGene cloningvGene expressionChapter one 第15页，本讲稿共34页3.proceeding 基因工程的过程由工程菌的设计构建和基因产物的生产两大部分组成。基因工程的过程由工程菌的设计构建和基因产物的生产两大部分组成。v从供体细胞中分离出基因组从供体细胞中分离出基因组DNA，用限制性内切酶分别将外源，用限制性内切酶分别将外源DNA(包含目的基因包含目的基因)和载体分子切和载体分子切开（简称切）开（简称切）v用用DNA连接酶将含有外源基因的连接酶将含有外源基因的DNA片断连接到载体分子上，形成片断连接

15、到载体分子上，形成DNA重组分子（简称接）重组分子（简称接）v借助于细胞转化手段将含有外源基因的借助于细胞转化手段将含有外源基因的DNA重组分子导入受体细胞中（简称转）重组分子导入受体细胞中（简称转）v短时间培养转化细胞，使短时间培养转化细胞，使DNA重组分子扩增或使其整合到受体细胞的基因组中（简称增）重组分子扩增或使其整合到受体细胞的基因组中（简称增）v筛选和鉴定转化细胞，获得使外源基因高效稳定表达的基因工程菌或细胞（简称检）筛选和鉴定转化细胞，获得使外源基因高效稳定表达的基因工程菌或细胞（简称检）v基因产物的表达与分离基因产物的表达与分离 基因工程的上游操作过程：切、接、转、增、检。基因工

16、程的上游操作过程：切、接、转、增、检。Chapter one 第16页，本讲稿共34页三、三、development of genetic engineeringvIn 1977，Genentech,inc.reports the production of the first human protein manufactured in a bacteria（E.coli）:somatostatin,a human growth hormone-releasing inhibitory factor(生长激素释放抑生长激素释放抑制素）制素）.vIn 1978,in the laboratory

17、 of Herbert Boyer at the University of California at San Francisco,a synthetic version of the human insulin（胰岛素）（胰岛素）gene was constructed and inserted into the bacterium Escheria coli.Chapter one 第17页，本讲稿共34页vIn 1979 John Baxter reported cloning the gene for human growth hormone(生长激素）生长激素）.vIn 1982

18、Genentech,Inc.received approval from the Food and Drug Administration to market genetically engineered human insulin,the first genetically engineered drug,a form of human insulin produced by bacteria.Chapter one 第18页，本讲稿共34页vIn 1981 scientists at Ohio University produced the first transgenic animals

19、 by transferring genes(growth hormone,生长激素生长激素)from other animals into mice.Chapter one 第19页，本讲稿共34页vIn 1985 genetically engineered plants resistant to insects,viruses,and bacteria were field tested for the first time.In 1986 the EPA(Environmental Protection Agency,美国环保署美国环保署)approved the release of

20、 the first genetically engineered crop,gene-altered tobacco plants.Chapter one 第20页，本讲稿共34页vIn 1985 the NIH(National Institutes of Health,全国全国卫生研究所卫生研究所)approved guidelines for performing experiments in gene therapy on humans.The first gene therapy takes place in 1990,on a four-year-old girl with an

21、 immune-system disorder.vThe Human Genome Project,the international effort to map all of the genes in the human body,was launched.1990 Formal launch of the international Human Genome Project.Chapter one 第21页，本讲稿共34页l人体一个基因组约人体一个基因组约6万个基因万个基因 l耗时耗时13年年l耗资耗资30亿美元亿美元l1Chapter one 第22页，本讲稿共34页 DNA conte

22、nt of the haploid(单倍体）单倍体）genome is related to the morphological complexity of lower eukaryotes,but varies extensively among the higher eukaryotes.The range of DNA values within a phylum is indicated by the shaded area.Chapter one 第23页，本讲稿共34页vIn 1997,Researchers at Scotlands Roslin Institute report

23、 that they have cloned a sheep-named Dolly-from the cell of an adult ewe（母羊）（母羊）.Chapter one 第24页，本讲稿共34页Chapter one 第25页，本讲稿共34页vIn 2002,A baby has been born through cloning,French scientist and member of the Raelian sect Brigitte Boisselier told AFP(法新社）法新社）.Chapter one Boisselier第26页，本讲稿共34页四、四、P

24、urposes for genetic engineering Genetic engineering、genetic modification or genetic manipulation(GM)All three terms mean the same thing,a laboratory technique used by scientists to change the genetic materials(DNAs)of living organisms,reshuffling of genes(重组基因重组基因)usually from one species to another

25、,for example:A gene from E.coli to tomato or from human to pig.Chapter one 第27页，本讲稿共34页v大规模生产生物制剂。如通过构建基因工程菌，使之大量大规模生产生物制剂。如通过构建基因工程菌，使之大量 产生产生具有应用价值的生物分子具有应用价值的生物分子酶制剂的生产、医药抗生素、维生酶制剂的生产、医药抗生素、维生素、氨基酸，研究肽疫苗等。素、氨基酸，研究肽疫苗等。-Genetic engineering medicines(药物药物)in therapeutics(治疗治疗)：insulin 胰岛素胰岛素 -inter

26、feron 干扰素干扰素 vaccines 疫疫 苗苗 -production of enzyme Chapter one 第28页，本讲稿共34页v设计构建新型物种。借助基因重组，基因定向突变以设计构建新型物种。借助基因重组，基因定向突变以及基因的人工合成技术，创造出具有新性状的物种。及基因的人工合成技术，创造出具有新性状的物种。如荧光鱼，黄金米，长着人耳的小鼠（干细胞）等。如荧光鱼，黄金米，长着人耳的小鼠（干细胞）等。-to increase resistance to disease or external damage,e.g.crops -to enable it to do som

27、ething it would not normally do Chapter one 第29页，本讲稿共34页Human DNA in a Goat CellThis goat contains a human gene that codes for a blood clotting agent.The blood clotting agent can be harvested in the goats milk.Transgenic Goat第30页，本讲稿共34页Chapter one 第31页，本讲稿共34页1.Create recombinant bacteria with desi

28、red gene.2.Allow the bacteria to“infect the plant cells.3.Desired gene is inserted into plant chromosomes.How to Create a Genetically Modified PlantChapter one 第32页，本讲稿共34页v破解生物的遗传信息资源，获得全部的生物基因组，破解生物的遗传信息资源，获得全部的生物基因组，为鉴定物种，研究生物进化，解开生物奥秘打下良好为鉴定物种，研究生物进化，解开生物奥秘打下良好的基础。的基础。Chapter one 第33页，本讲稿共34页Chapter one 第34页，本讲稿共34页