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Cloning, Gene Expression, and Protein Purification: Experimental Procedures and Process Rationale [Paperback]

Charles Hardin (Author), Jennifer Edwards (Author), Andrew Riell (Author), David Presutti (Author), William Miller (Author), Dominique Robertson (Author)

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Book Description

March 1, 2001 0195132947 978-0195132946 1
On the forefront of modern scientific innovation, Cloning, Gene Expression and Protein Purification: Experimental Procedures and Process Rationale effectively doubles as a laboratory manual for students and a reference book for professional researchers. Designed for advanced undergraduate and beginning graduate students in molecular biology, this unique combination lecture/laboratory resource presents detailed protocols for the multi-step process involved in isolating a gene, cloning and characterizing it, expressing its encoded protein, and purifying and characterizing the protein's basic physical properties. This manageable volume includes both theoretical background and practical procedures and is structured around twenty experiments that demonstrate how to prepare, manipulate, and analyze plasmids, produce fusion proteins in bacteria, and purify these proteins based on unique chemical properties or substrate affinities. The book describes advanced topics such as the use of antibodies and the techniques developed to transform their structures, as well as combinatorial approaches designed to manipulate the structure and functions of proteins and nucleic acids. Supplemental literature provides a variety of theoretical explanations encouraging a more intuitive understanding of the experimental mechanisms and behaviors of the chemical participants, while also giving students the tools needed to become "capable proactive researchers." Features:
� Emphasizes electrophoresis, Southern and Western blotting, and combinatorial techniques
� Defines clear reaction mechanisms; stipulates the functions of reagents; and helps students think about the precise consequences of solution and procedural manipulations
� Discusses fluorophores, and solvent effects on protein structure
� Characterizes plasmids, cDNAs, and antibody probes (available from ATCC) in research literature
� Includes carefully selected primary source research literature and articles from current vendor literature
� Contains a glossary of unfamiliar phrases and jargon; important summary statements and conclusions are italicized
� Provides an alphabetized list of common reagents for rapid reference
� Offers an extensive index of concepts and terms
� Categorizes helpful and distinctive information into five types of supplemental literature: Innovation/ Insight, Theory/Principle, Process Rationale, Vendor Literature, and Alternative Approaches

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About the Author


Charles Hardin is an Associate Professor of Biochemistry at North Carolina State University. Jennifer Pinczes recently completed her Master's of Science in biochemistry at North Carolina State University. Andrew Riell is a computer specialist and consultant with NetAspects, Inc. William Miller is William Neals Reynolds Professor of Biochemistry at North Carolina State University. David Presutti is an Adjunct Visiting Professor of Biochemistry at North Carolina State University. Dominique Robertson is an Associate Professor of Botany at North Carolina State University.

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Inside This Book (learn more)
First Sentence:
This set of experiments is organized as a set of interconnected processes. Read the first page
Key Phrases - Statistically Improbable Phrases (SIPs): (learn more)
dominant folding force, floating tube rack, filtrate collector, fusion protein expression vector, pure macromolecules, plunger knob, nonpolar solvation, ligase reaction buffer, hexanucleotide mixture, bicinchonic acid, glycinate ions, glass pellet, glass concentrate, detection buffer, working reagent, labeling grade, biotinylated oligo, intrinsic propensities, intrachain hydrogen bonds, substrate mix, lac repressor protein, sorbent extraction, reduction buffer, nonpolar contacts, coding fragment
Key Phrases - Capitalized Phrases (CAPs): (learn more)
New York, Experimental Outline, Required Materials, Cold Spring Harbor Laboratory, Methods Enzymol, Protein Chem, Nucleic Acids Res, John Wiley, Additional Report Requirements, Color Substrate Solution, Academic Press, Boehringer Mannheim, Genius System, Protein Eng, Coomassie Brilliant Blue, Current Protocols, Laboratory Manual, Molecular Cloning, Scripps Research Institute, University of California, Prelab Preparations, Additional Reading, Purification Kit, San Diego, Storage Handling
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