Phage display : a practical approach / edited by Tim Clackson, Henry B. Lowman.

This new book aims to enable researchers to design and undertake all aspects of a phage display project, from designing an experimental strategy and constructing a library to performing selections and analyzing the results. All of the protocols and chapters are extensively cross-referenced, allowing...

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Bibliographic Details
Other Authors: Clackson, Tim, Lowman, Henry B.
Format: eBook
Language:English
Published: Oxford ; New York : Oxford University Press, ©2004.
Series:Practical approach series ; 266.
Subjects:
Online Access:Click for online access

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245 0 0 |a Phage display :  |b a practical approach /  |c edited by Tim Clackson, Henry B. Lowman. 
260 |a Oxford ;  |a New York :  |b Oxford University Press,  |c ©2004. 
300 |a 1 online resource (xxiv, 332 pages) :  |b illustrations 
336 |a text  |b txt  |2 rdacontent 
337 |a computer  |b c  |2 rdamedia 
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490 1 |a Practical approach series ;  |v no. 266 
504 |a Includes bibliographical references and index. 
588 0 |a Print version record. 
505 0 |a Cover -- Contents -- Protocol list -- List of abbreviations -- List of contributors -- 1 Introduction to phage biology and phage display -- 1 Introduction -- 2 Biology of filamentous phage -- 2.1 Introduction -- 2.2 Structure of the phage particle -- 2.3 Infection -- 2.4 Replication -- 2.5 Genes and gene expression -- 2.6 Physiology of phage assembly -- 2.7 The mechanics of phage assembly -- 3 Coat proteins used for display -- 3.1 pVIII -- 3.2 pIll -- 4 Starting a phage display project -- 4.1 Feasibility of display -- 4.2 Phage or phagemid vector? -- 4.3 Polyvalent or monovalent display? -- 4.4 Helper phage -- 4.5 General protocols for phage preparation and quantitation -- 5 General principles of a phage display project -- 5.1 Making a library -- 5.2 Selection -- 5.3 Analysis of clones -- 6 Common problems -- 6.1 Library quality -- 6.2 Expression editing -- 6.3 Over-selection -- 7 Alternative display systems -- 8 Commercial sources of phage display libraries and kits -- References -- 2 Constructing phage display libraries by oligonucleotide-directed mutagenesis -- 1 Introduction -- 2 Considerations for library design -- 2.1 Site-directed mutagenesis -- 2.2 Degenerate codon design -- 2.3 Theoretical versus actual diversity -- 3 Oligonucleotide-directed mutagenesis -- 3.1 Oligonucleotide-directed mutagenesis versus cassette mutagenesis -- 3.2 The chemistry and biology of oligonucleotide-directed mutagenesis -- 3.3 Construction of an inactive template -- 4 Library construction and storage -- 4.1 Preparation of single-stranded DNA template -- 4.2 In vitro synthesis of heteroduplex CCC-dsDNA -- 4.3 E.coli electroporation and production of library phage -- 4.4 Library storage and reinfection -- 5 Biological reagents -- References -- 3 In vitro DNA recombination -- 1 Introduction -- 2 Background to in vitro DNA recombination -- 2.1 Use of in vitro DNA recombination in directed evolution -- 2.2 Applications of in vitro DNA recombination -- 2.3 Recombination statistics -- 3 Methods for in vitro DNA recombination -- 3.1 Stemmer method -- 3.2 Random DNA fragmentation with endonuclease V from E. coli -- 3.3 Random priming recombination -- 3.4 Staggered Extension Process (StEP) -- 3.5 In vitro heteroduplex formation and in vivo repair (heteroduplex recombination) -- 3.6 Choice of recombination method -- 3.7 Removal of background -- 3.8 Technical tips -- References -- 4 Phage selection strategies for improved affinity and specificity of proteins and peptides -- 1 Introduction -- 2 Vector considerations -- 2.1 Monovalent and polyvalent phage display -- 2.2 Confirming display -- 2.3 Protein expression from phagemid vectors -- 2.4 Vector construction and phagemid preparation -- 3 Library design -- 3.1 Hard randomization -- 3.2 Soft randomization -- 4 Target presentation -- 4.1 Direct immobilization -- 4.2 Solution binding -- 4.3 Blocking -- 4.4 Pilot selection -- 5 Selection -- 5.1 Binding buffer considerations -- 5.2 Stringency of selection -- 5.3 Competitive selection -- 5.4 Elution of bound phage -- 5.5 Amplification -- 5.6 Monitori. 
520 |a This new book aims to enable researchers to design and undertake all aspects of a phage display project, from designing an experimental strategy and constructing a library to performing selections and analyzing the results. All of the protocols and chapters are extensively cross-referenced, allowing readers to move beyond the specific examples provided in order to customize the procedures for their own protein or selection system of interest. Phage Display is an up-to-date, . comprehensive and integrated experimental guide to the technique, which is essential reading for anyone currently using. 
650 0 |a Bacteriophages. 
650 0 |a Viral proteins. 
650 0 |a Microbial biotechnology. 
650 7 |a SCIENCE  |x Life Sciences  |x Biology.  |2 bisacsh 
650 7 |a SCIENCE  |x Life Sciences  |x Microbiology.  |2 bisacsh 
650 7 |a Bacteriophages  |2 fast 
650 7 |a Microbial biotechnology  |2 fast 
650 7 |a Viral proteins  |2 fast 
700 1 |a Clackson, Tim. 
700 1 |a Lowman, Henry B. 
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830 0 |a Practical approach series ;  |v 266. 
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