Genetic Engineering 基因工程

1. GeneticEngineering基因工程 Jacky Sun, Ph.D. Molecular Biology

2. Syllabus课程提纲 Course课程: Genetic Engineering Time: Spring Semester, 2013 References参考:Gene VIII (基因八[美] 莱威·本杰明) Essentials要素of Current当代Protocol in Molecular Biology (Roger Brent, 2003) Molecular Cloning: A Laboratory Manual指南(Joseph Sambrook, 2001) Instructor: Guopeng Sun, Ph.D. Genetic Engineering

3. Objective1目标 • Learn and understand some important concepts概念and definitions定义of genetic engineering. • Learn and understand some basic methods used in recombinant重组DNA manipulation操作 Genetic Engineering

4. Objective2 • Learn and understand what we can do to use the theories and methods of genetic engineering. • Learn and understand the standard operation of the experiments that are important methods in genetic engineering. Genetic Engineering

5. Grading评分 • Attendance出勤: 10% • Experimental techniques：10% • Two Take-home Tests： 10% • Class participation参与： 10% • Final exam: 60% Genetic Engineering

6. Schedule进度表 Genetic Engineering

7. Chapter 1: Introduction • Definition of Genetic Engineering • Significance意义and Concerns • History and Development • Mainstayof Genetic Manipulation • Common Techniques in Genetic Engineering • Concept Map Genetic Engineering

8. 1. Definition Genetic Engineering: tools and techniques used to isolate分离one or more genes from an organism, modify修饰the material obtained, and transfer转移it into another organism Recombinant DNA technology: techniques of gene cloning. Recombinant DNA: the molecule formed by joining连接a DNA of interest to vector DNA Genetic Engineering

9. Isolation Interest Gene PCR/Cloning Target sequence Vector Enzyme sites Genetic Engineering

10. Genetic Engineering

11. 2. Significance and Concerns • Significance • Concerns Genetic Engineering

12. Significance • Basic research on gene structure and function • Production of useful proteins by novel新颖的methods • Generation of transgenic plants and animals • Medical diagnosis诊断and treatment. Genetic Engineering

13. 1.Basic research on gene structure and function Genetic Engineering

14. 2.Production of useful proteins by novel methods eucaryotic prokaryotic Genetic Engineering

15. 3.Generation of transgenic plants and animals Adversity抗逆基因gene Fluorescence荧光gene Genetic Engineering

16. Concerns担忧 1. The ethical伦理的problems Genetic Engineering

17. Concerns担忧 2. The problems of Genetically modified foods Some research confirmed that the GM food could cause infertility不孕in mouse. However, there was no evidencethat could happened to human. Genetic Engineering

18. 3. History and Early Development • Three Main Eras时代of Development of Genetics • Early Development of Gene Cloning Genetic Engineering

19. Three Main Eras of Development of Genetics1 • Rediscovery of Gregor Mendel‘s work in early 1900, elucidation阐明of the inheritance遗传principles原理and genetic mapping in the next 40 years. Genetic Engineering

20. Three Main Eras of Development of Genetics2 • Establishment确立of microbial genetics in mid- 1940s, role of DNA as the genetic material was confirmed证实, understanding the mechanisms机制of gene transfer between bacteria Genetic Engineering

21. Three Main Eras时代of Development of Genetics3 • Discovery of the structure of DNA by James Watson and Francis Crick in 1953 leads genetics to molecular level, establishment of the complete genetic code in 1966 - the stage was now set for the appearance出现of the new genetics. Genetic Engineering

22. The Nobel Prize in Physiology or Medicine 1962 Francis H. C. Crick James D. Watson Maurice H. F. Wilkins "for their discoveries concerning the molecular structure of nucleic acids and its significance for information transfer in living material" Genetic Engineering

23. Early Development of Gene Cloning • Molecular glue胶水- DNA ligase isolated in 1967 • Molecular scissors剪刀- the first restriction限制性enzyme isolated in 1970 • First recombinant DNA molecules分子generated at Stanford University in 1972 • Joining DNA fragments片段to the plasmid质粒pSC101 in 1973 Genetic Engineering

24. 4. Mainstay中流砥柱of Genetic Manipulation (Discussion in class) • Target目标Gene cloning: Single DNA fragment isolation; the genomic or cDNA library文库construction建造and screening筛选; PCR • Recombinant DNA construction or modification修饰in vitro体外:restriction enzymes, modifying enzymes, vectors; PCR • DNA amplification扩增in vivo体内:transformation转化and selection in vitro; Research and application: recombinant protein expression, transgenic, cloning, medical diagnosis诊断, gene therapy Genetic Engineering

25. Genetic Engineering

26. 5. Common Techniques • Southern Hybridization杂交 • Northern Hybridization • PCR+ • Restriction digestion + • DNAsequencing • DNA cloning and Transformation + • Mutagenesis in vitro+ Genetic Engineering

27. Aragrose Gel Electrophoresis to Separate DNA Agarose is an inert惰性carbohydrate碳水化合物isolated from seaweeds. DNA molecules in the gel can be visualized直观by staining染色with EB and observe under UV light. Genetic Engineering

28. DNA Fragments Visualized Under UV Light Genetic Engineering

29. Southern Hybridization1 Genetic Engineering

30. Southern Hybridization2 Genetic Engineering

31. Northern Hybridization Genetic Engineering

32. Restriction Digestion and Ligation Genetic Engineering

33. Cloning by Plasmid Genetic Engineering

34. Transformation + Plasmid DNA Bacteria picks up a piece of DNA Genetic Engineering

35. PCR Reaction Genetic Engineering

36. Genomic PCR 1.5 kb 0.2 kb 0.1 kb Genetic Engineering

37. DNA Sequencing Genetic Engineering

38. Automated DNA sequencing Genetic Engineering

39. Mutagenesis in vitro Genetic Engineering

40. 6. Concept Map Genetic Engineering