http://spxy.zjgsu.edu.cn 《基因工程原理和技术（英）》教学大纲 (Genetic Engineering) 一、基本信息 课程代码：1040022 学 分： 2 总 课 时：32 课程性质：专业选修课 适用专业：生物工程 先修课程：生物化学，微生物学，分子生物学 二、本课程教学目的和任务 基因工程原理与技术是为生物工程专业开设的一门专业选修课。通过课程学习，使学生 更进一步了解生物技术领域中的前沿技术，并扎实专业理论知识。 三、教学方法与手段 多媒体教学。 四、教学内容及要求 Chapter 1 Gene manipulation: an all-embracing technique To have a good grasp of the concepts of gene manipulation and content. To be familiar with the content of sequence analysis, in vivo biochemistry, the new medicine, biotechnology, the central role of E. coli. To have an idea of the historical development of genetic engineering. Key points：The concepts of gene manipulation and content. Chapter 2 Basic techniques To have a good grasp of the techniques of agarose gel electrophoresis, nucleic acid blotting, transformation of E. coli and PCR. To be familiar with the content of real-time quantitative PCR. To know the content of transformation of other organisms. Key points：transformation of E. coli and PCR. Difficult points：real-time quantitative PCR. Chapter 3 Cutting and joining DNA molecules To have a good grasp of the joining DNA molecules. To be familiar with the content of host-controlled restriction and modification. To know the content of the Dam and Dcm methylases of E. coli , the importance of eliminating restriction systems in E. coli strains used as hosts for recombinant molecules. Key points：the joining DNA molecules. Difficult points： host-controlled restriction and modification, and the Dam and Dcm methylases of E. coli. http://spxy.zjgsu.edu.cn Chapter 4 Basic biology of plasmid and phage vectors To have a good grasp of the plasmid biology and simple plasmid vectors, bacteriophage λ. To be familiar with the content of the purification of plasmid DNA. To know the content of DNA cloning with single-stranded DNA vecters. Key points：the plasmid biology and simple plasmid vectors, bacteriophage λ. Difficult points：bacteriophage λ. Chapter 5 Cosmids, phasmids and other advanced vectors To have a good grasp of vectors for cloning large fragments of DNA. To be familiar with the content of specialist-purpose vectors. To know the content of putting it all together: vectors with combination of features. Key points：vectors for cloning large fragments of DNA. Difficult points：vectors with combination of features. Chapter 6 Cloning strategies To have a good grasp of cloning genomic DNA. To be familiar with the cDNA cloning, screening strategies. To know the content of difference cloning. Key points：cloning genomic DNA. Difficult points：screening strategies. Chapter 7 Sequencing and mutagenesis To have a good grasp of basic DNA sequencing, whole-genome sequencing. To be familiar with analyzing sequencing data. To know the content of changing genes:site-directed mutagensis. Key points：basic DNA sequencing, whole-genome sequencing. Difficult points：changing genes:site-directed mutagensis. Chapter 8 Cloning in bacteria other than Escherichia coli To have a good grasp of introducing DNA into bacterial cells. To be familiar with cloning in Gram-negative bacteria other than E. coli, cloning in Gram-positive bacteria. To know the content of cloning in streptomycetes. Key points：introducing DNA into bacterial cells. Difficult points：cloning in streptomycetes. Chapter 9 Cloning in Saccharomyces cerevisiae and other fungi To have a good grasp of introducing DNA into fungi, expression of cloned genes. To be familiar with yeast surface display. To know the content of identifying genes encoding particular cellular activities, determining functions associated with particular genes. Key points：introducing DNA into fungi, expression of cloned genes. Difficult points：yeast surface display. Chapter 10 Gene transfer to animal cells To have a good grasp of DNA-mediated transformation, gene transfer by viral transduction. To be familiar with expression systems for animal cell. http://spxy.zjgsu.edu.cn Key points：DNA-mediated transformation. Difficult points：gene transfer by viral transduction. Chapter 11 Genetic manipulation of animals To have a good grasp of genetic manipulation of mammals. To be familiar with DNA transfer to other vertebrates, DNA transfer to invertebrates. Key points：genetic manipulation of mammals. Difficult points：genetic manipulation of mammals. Chapter 12 Gene transfer to plants To have a good grasp of Agrobacterium-mediated transformation. To be familiar with chloroplast transformation, plant viruses as vectors. To know the content of direct DNA transfer to plant. Key points：Agrobacterium -mediated transformation. Difficult points：chloroplast transformation. Chapter 13 Advances in transgenic technology To have a good grasp of further transgenic strategies for gene inhibition, inducible expression systems. To be familiar with applications of site-specific recombination. To know the content of transgenic technology for functional genomics. Key points：further transgenic strategies for gene inhibition, applications of site-specific recombination. Difficult points：applications of site-specific recombination. Chapter 14 Application of recombinant DNA tecgnology To have a good grasp of nucleic acid sequences as diagnostic tools, protein engineering and metabolic engineering. To be familiar with plant breeding in the twenty-first century and new drugs and new therapies for genetic diseases. Key points： nucleic acid sequences as diagnostic tools, protein engineering and metabolic engineering. 五、教学时数分配表 Chapter Content 参考时数 1 Gene manipulation: an all-embracing technique 2 2 Basic techniques 2 3 Cutting and joining DNA molecules 2 4 Basic biology of plasmid and phage vectors 2 5 Cosmids, phasmids and other advanced vectors 2 6 Cloning strategies 2 7 Sequencing and mutagenesis 2 8 Cloning in bacteria other than Escherichia coli 4 9 Cloning in Saccharomyces cerevisiae and other fungi 4 http://spxy.zjgsu.edu.cn 10 Gene transfer to animal cells 2 11 Genetic manipulation of animals 2 12 Gene transfer to plants 2 13 Advances in transgenic technology 2 14 Application of recombinant DNA tecgnology 2 Total 32 六、考核方式及成绩评定标准 考核方式：闭卷 成绩评定标准： 平时成绩占 30%；期末成绩占 70%。成绩评定为百分制。总成绩=平时成 绩×30%+期末成绩×70%。 七、教材及主要参考书 教材 ： Principles of Gene Manipulation, Pimrose S, et al. Sixth Edition. Higher Education Press, 2002 参考书目： [1] From gene to genomes: Concepts and applications of DNA technology, Dale JW, et al. First Edition. Science Press,2004 [2] 吴乃虎《基因工程原理》 科学出版社 第一版1999 年 [3] 朱玉贤 主编 《现代分子生物学》 高等教育出版社 第二版 2002 年John Wiley, [4] 张惠展 主编 《基因工程概论》 华东理工大学出版社 第一版 1999 年 执笔人:顾 青