[其它]CHapter1-2.ppt

Gene Engineering References: 1、Gene manipulation 6th Ed. Sandy Primrose, et al. Higher Education Press, 2002 2、基因操作原理， 瞿礼佳等译，高等教育出版社，2002 3、基因工程原理（第二版），吴乃虎著，科学出版社，1998 4、Gene cloning: a laboratory mannual(3rd Ed.) Sambrook J and Russel DW , Cold Spring Harbor Laboratory Press,2001 CHAPTER 1 Gene manipulation: an all-embracing technique Introduction 1. gene manupulation (gene engineering): the formation of new combinations of heritable material by the insertion of nucleic acid molecules, produced by whatever means outside the cell, into any virus, bacterial plasmid or other vector system so as to allow their incorporation into a host organism in which they do not naturally occur but in which they are capable of continued propagation. 2.in vivo: alive creature or human body 3.in vitro: outside of creature or human body Sequence analysis 1. Methods: F. Sanger (dideoxy chain terminating method), Maxam-Gilbert (chemical degradation method); Manually sequencing, Automatically sequencing 2. Applications: 1)? Open reading frame (ORF) 2) cis-acting elements: e.g. Promoter, Terminator, Enhancer 3)?Prediction of DNA structures: e.g. Z-DNA, Palindrome 4)?Prediction of protein structure: primary structure, secondary structure, three-dimension structure In vivo biochemistry 1.? cell components studied in vitro: e.g. enzyme- purification, activity study 2.? cell components studied in vivo: e.g. mutants 1)?occurred naturally 2)?treatment by physical and chemical methods: e.g. UV, EB 3)?gene manipulation: gene knock-out, gene knock-in The new medicine 1.?protein drugs: e.g. HGH 2.?Vaccines: e.g. HBV 3. Small molecules( plants, microbes) 1) over-express genes relating biosynthetic pathway 2)?shut down gene expression to reduce the intermediates towards the by-products 4.?Diagnostic nucleic acid: pathogens, genetic diseases 5.? Therapeutic nucleic acid: 1)? repair of gene defects 2)?anti-sense nucleic acid: down-regulated gene expression Biotechnology: the new