Gene Cloning DNA Analysis An Introduction.pdf
《Gene Cloning DNA Analysis An Introduction.pdf》由会员分享,可在线阅读,更多相关《Gene Cloning DNA Analysis An Introduction.pdf(338页珍藏版)》请在沃文网上搜索。
1、9781405181730_1_pre.qxd 1/11/10 10:23 Page ivGENECLONINGAND DNAANALYSIS9781405181730_1_pre.qxd 1/11/10 10:23 Page i9781405181730_1_pre.qxd 1/11/10 10:23 Page iiGENECLONINGAND DNAANALYSISAn IntroductionT.A.BROWNFaculty of Life SciencesUniversity of ManchesterManchesterSixth EditionA John Wiley&Sons,L
2、td.,Publication9781405181730_1_pre.qxd 1/11/10 10:23 Page iiiThis edition first published 2010,2010,2006 by T.A.BrownFirst,second and third editions published by Chapman&Hall 1986,1990,1995Fourth and fifth editions published by Blackwell Publishing Ltd 2001,2006Blackwell Publishing was acquired by J
3、ohn Wiley&Sons in February 2007.Blackwells publishing program has been merged with Wileys global Scientific,Technical and Medical business to form Wiley-Blackwell.Registered office:John Wiley&Sons Ltd,The Atrium,Southern Gate,Chichester,West Sussex,PO19 8SQ,UKEditorial offices:9600 Garsington Road,O
4、xford,OX4 2DQ,UKThe Atrium,Southern Gate,Chichester,West Sussex,PO19 8SQ,UK111 River Street,Hoboken,NJ 07030-5774,USAFor details of our global editorial offices,for customer services and for information about how to apply for permission to reuse the copyright material in this book please see our web
5、site at right of the author to be identified as the author of this work has been asserted in accordance with the Copyright,Designs and Patents Act 1988.All rights reserved.No part of this publication may be reproduced,stored in a retrieval system,or transmitted,in any form or by any means,electronic
6、,mechanical,photocopying,recording or otherwise,except as permitted by the UK Copyright,Designs and Patents Act 1988,without the prior permission of the publisher.Wiley also publishes its books in a variety of electronic formats.Some content that appears in print may not be available in electronic b
7、ooks.Designations used by companies to distinguish their products are often claimed as trademarks.All brand names and product names used in this book are trade names,service marks,trademarks or registered trademarks of their respective owners.The publisher is not associated with any productor vendor
8、 mentioned in this book.This publication is designed to provide accurate and authoritativeinformation in regard to the subject matter covered.It is sold on the understanding that thepublisher is not engaged in rendering professional services.If professional advice or other expertassistance is requir
9、ed,the services of a competent professional should be sought.Library of Congress Cataloguing-in-Publication DataBrown,T.A.(Terence A.)Gene cloning and DNA analysis:an introduction/T.A.Brown.6th ed.p.cm.ISBN 978-1-4051-8173-0(pbk.:alk.paper)ISBN 978-1-4443-3407-4(hbk.:alk.paper)1.Molecular cloning.2.
10、Nucleotide sequence.3.DNAAnalysis.I.Title.QH442.2.B76 2010572.8633dc222009038739ISBN:9781405181730(paperback)and 9781444334074(hardback)A catalog record for this book is available from the British Library.Set in 10/12pt Classical Garamond by Graphicraft Limited,Hong KongPrinted in Malaysia1201097814
11、05181730_1_pre.qxd 1/11/10 10:23 Page ivPreface to the Sixth Edition xviPart I The Basic Principles of Gene Cloning and DNA Analysis11Why Gene Cloning and DNA Analysis are Important32Vectors for Gene Cloning:Plasmids and Bacteriophages133Purification of DNA from Living Cells254Manipulation of Purifi
12、ed DNA 455Introduction of DNA into Living Cells726Cloning Vectors for E.coli887Cloning Vectors for Eukaryotes1058How to Obtain a Clone of a Specific Gene1269The Polymerase Chain Reaction147Part II The Applications of Gene Cloning and DNA Analysis in Research16310Sequencing Genes and Genomes16511Stud
13、ying Gene Expression and Function18512Studying Genomes207Part III The Applications of Gene Cloning and DNA Analysis in Biotechnology22313Production of Protein from Cloned Genes22514Gene Cloning and DNA Analysis in Medicine24515Gene Cloning and DNA Analysis in Agriculture26416Gene Cloning and DNA Ana
14、lysis in Forensic Science and Archaeology282Glossary298Index312Companion website available at CONTENTSBrief Contents9781405181730_1_pre.qxd 1/11/10 10:23 Page v9781405181730_1_pre.qxd 1/11/10 10:23 Page viPreface to the Sixth EditionxviPart I The Basic Principles of Gene Cloning and DNA Analysis11 W
15、hy Gene Cloning and DNA Analysis are Important31.1The early development of genetics31.2The advent of gene cloning and the polymerase chain reaction41.3What is gene cloning?51.4What is PCR?61.5Why gene cloning and PCR are so important71.5.1Obtaining a pure sample of a gene by cloning71.5.2PCR can als
16、o be used to purify a gene91.6How to find your way through this book112 Vectors for Gene Cloning:Plasmids andBacteriophages132.1Plasmids132.1.1Size and copy number152.1.2Conjugation and compatibility162.1.3Plasmid classification162.1.4Plasmids in organisms other than bacteria172.2Bacteriophages172.2
17、.1The phage infection cycle182.2.2Lysogenic phages19Gene organization in the 2 DNA molecule19The linear and circular forms of 2 DNA19M13a filamentous phage222.2.3Viruses as cloning vectors for other organisms24viiCONTENTSContents9781405181730_1_pre.qxd 1/11/10 10:23 Page vii3 Purification of DNA fro
18、m Living Cells253.1Preparation of total cell DNA253.1.1Growing and harvesting a bacterial culture263.1.2Preparation of a cell extract283.1.3Purification of DNA from a cell extract29Removing contaminants by organic extraction and enzymedigestion29Using ion-exchange chromatography to purify DNA from a
19、 cellextract303.1.4Concentration of DNA samples303.1.5Measurement of DNA concentration313.1.6Other methods for the preparation of total cell DNA323.2Preparation of plasmid DNA333.2.1Separation on the basis of size353.2.2Separation on the basis of conformation36Alkaline denaturation36Ethidium bromide
20、caesium chloride density gradientcentrifugation363.2.3Plasmid amplification393.3Preparation of bacteriophage DNA393.3.1Growth of cultures to obtain a high 2 titer403.3.2Preparation of non-lysogenic 2 phages403.3.3Collection of phages from an infected culture423.3.4Purification of DNA from 2 phage pa
21、rticles423.3.5Purification of M13 DNA causes few problems434 Manipulation of Purified DNA454.1The range of DNA manipulative enzymes464.1.1Nucleases464.1.2Ligases474.1.3Polymerases484.1.4DNA modifying enzymes494.2Enzymes for cutting DNArestriction endonucleases504.2.1The discovery and function of res
22、triction endonucleases514.2.2Type II restriction endonucleases cut DNA at specific nucleotidesequences524.2.3Blunt ends and sticky ends534.2.4The frequency of recognition sequences in a DNA molecule534.2.5Performing a restriction digest in the laboratory544.2.6Analysing the result of restriction end
23、onuclease cleavage56Separation of molecules by gel electrophoresis57Visualizing DNA molecules in an agarose gel584.2.7Estimation of the sizes of DNA molecules584.2.8Mapping the positions of different restriction sites in a DNAmolecule594.2.9Special gel electrophoresis methods for separating largermo
24、lecules60Contentsviii9781405181730_1_pre.qxd 1/11/10 10:23 Page viii4.3Ligationjoining DNA molecules together634.3.1The mode of action of DNA ligase634.3.2Sticky ends increase the efficiency of ligation644.3.3Putting sticky ends onto a blunt-ended molecule64Linkers64Adaptors65Producing sticky ends b
25、y homopolymer tailing674.3.4Blunt end ligation with a DNA topoisomerase695 Introduction of DNA into Living Cells725.1Transformationthe uptake of DNA by bacterial cells745.1.1Not all species of bacteria are equally efficient at DNA uptake745.1.2Preparation of competent E.coli cells755.1.3Selection fo
- 1.请仔细阅读文档,确保文档完整性,对于不预览、不比对内容而直接下载带来的问题本站不予受理。
- 2.下载的文档,不会出现我们的网址水印。
- 3、该文档所得收入(下载+内容+预览)归上传者、原创作者;如果您是本文档原作者,请点此认领!既往收益都归您。
下载文档到电脑,查找使用更方便
30 积分
下载 | 加入VIP,下载更划算! |
- 配套讲稿:
如PPT文件的首页显示word图标,表示该PPT已包含配套word讲稿。双击word图标可打开word文档。
- 特殊限制:
部分文档作品中含有的国旗、国徽等图片,仅作为作品整体效果示例展示,禁止商用。设计者仅对作品中独创性部分享有著作权。
- 关 键 词:
- Gene Cloning DNA Analysis An Introduction