Experiments in Molecular Biology
Biochemical Applications
Published on 4. March 1997
Book
Paperback/Softback
256 pages
978-0-12-147370-9 (ISBN)
Description
Experiments in Molecular Biology provides a thorough introduction to recombinant DNA methods used in molecular biology and nucleic acid biochemistry. This unique laboratory manual is particularly appropriate for courses in molecular cloning, molecular genetics techniques, molecular biology techniques, recombinant DNA techniques, bacterial genetics techniques, and genetic engineering. Included is an especially helpful section to aid new instructors in avoiding potential pitfalls of specific experiments.
More details
Language
English
Place of publication
San Diego
United States
Publishing group
Elsevier Science Publishing Co Inc
Target group
College/higher education
Dimensions
Height: 279 mm
Width: 216 mm
Weight
750 gr
ISBN-13
978-0-12-147370-9 (9780121473709)
Copyright in bibliographic data and cover images is held by Nielsen Book Services Limited or by the publishers or by their respective licensors: all rights reserved.
Schweitzer Classification
Persons
Dr. Burton has been a professor at Michigan State University since 1987 and is also Director of Undergraduate Education in the Department of Biochemistry and Molecular Biology, as well as Principle Investigator of the Burton Lab at MSU. His research is on RNA polymerase mechanisms and ancient evolution of transcription and translation systems, on which he has published several papers. He is interested in the genesis, divergence, and complexity of life on Earth, told in core protein and DNA sequences.
Content
Introduction to the Laboratory Course on Biochemistry and Molecular Biological Methods:
An Essay on Science and Scientists.
Introduction to the Course.
Principles of Molecular Genetics and Nucleic Acid Biochemistry:
Bacteria: Composition, Organization, and Growth.
Methods to Measure the Growth of a Bacterial Culture.
Growth of Cultures.
Basic Techniques for Bacterial Genetics.
Bacteriophage Lambda.
Biology.
The Decision: Lysis versus Lysogeny.
The Regulators.
Transcriptional Antiterminators.
Lytic Growth.
Lysogenic Growth.
Induction of the Prophage.
Regulation of Lambda int Gene Expression by Regulating mRNA Stability.
Lambda as a Transducing Phage.
Plating and Counting Phage.
Maintenance of Lambda Stocks.
Lambda ZAPII/topA.
Plasmid Vectors.
pBR322.
Transferring Plasmid DNA into Bacterial Cells.
Storage of Plasmid-Containing Escherichia coli Strains and of Plasmid DNA.
Expression Vectors.
Translation Signals in Bacterial Cells and Human Cells.
Expression Vectors Based on Transcription from Lambda PL.
Expression Vectors Based on the Lactose Promoter.
Lactose Operon Biology.
Regulation of Transcription from Plac.
An Improved Plac for Recombinant Systems.
An Improved Inducer.
The Expression Vector pTrc99A.
Expression Systems Using Alternate RNA Polymerases.
Topoisomerases.
Topological Forms of a Circular Duplex DNA.
Mechanism of Action of DNA Gyrase.
Mechanism of Action of Topoisomerase I.
Formation of Catenated DNAs.
Enzymatic Methods to Measure Topoisomerase I Activity.
Inserting DNA into a Cloning Vector.
Restriction Endonucleases.
DNA Ligases.
Phosphatase Treatment of Cloning Vectors.
Inter- and Intramolecular Joining of DNA Ends.
Cloning Strategies.
Insertional Inactivation.
Directional Cloning.
Replacement Cloning.
Construction of Recombinant DNA Libraries.
Lambda Libraries.
Lambda as a Cloning Vector.
Lambda ZAP II, an Insertion Vector.
cDNA Synthesis.
Replacement Vectors.
Purposes for Genomic and cDNA Cloning.
Methods for Detection of Clones.
Hybridization Methods.
Plaque Hybridization.
Colony Hybridization.
Histochemical Detection of Beta-Galactosidase.
Alpha Complementation.
pBluescript II SK-.
Antibody Screening Methods.
The BCIP/NBT Color Development Reaction for Antibody Detection.
M13 as a Cloning Vector.
Cloning Vectors of M13 and the Role of the lac Operon.
Comparison of M13 and Lambda.
The Life Cycle of M13 and Its Mode of Replication.
Phagemids.
DNA Sequencing and in Vitro Mutagenesis.
Enzymatic and Chemical Methods of DNA Sequence Analysis.
The Enzymatic Method.
The Chemical Method.
Laboratory Experiments.
Appendices.
Subject Index.
An Essay on Science and Scientists.
Introduction to the Course.
Principles of Molecular Genetics and Nucleic Acid Biochemistry:
Bacteria: Composition, Organization, and Growth.
Methods to Measure the Growth of a Bacterial Culture.
Growth of Cultures.
Basic Techniques for Bacterial Genetics.
Bacteriophage Lambda.
Biology.
The Decision: Lysis versus Lysogeny.
The Regulators.
Transcriptional Antiterminators.
Lytic Growth.
Lysogenic Growth.
Induction of the Prophage.
Regulation of Lambda int Gene Expression by Regulating mRNA Stability.
Lambda as a Transducing Phage.
Plating and Counting Phage.
Maintenance of Lambda Stocks.
Lambda ZAPII/topA.
Plasmid Vectors.
pBR322.
Transferring Plasmid DNA into Bacterial Cells.
Storage of Plasmid-Containing Escherichia coli Strains and of Plasmid DNA.
Expression Vectors.
Translation Signals in Bacterial Cells and Human Cells.
Expression Vectors Based on Transcription from Lambda PL.
Expression Vectors Based on the Lactose Promoter.
Lactose Operon Biology.
Regulation of Transcription from Plac.
An Improved Plac for Recombinant Systems.
An Improved Inducer.
The Expression Vector pTrc99A.
Expression Systems Using Alternate RNA Polymerases.
Topoisomerases.
Topological Forms of a Circular Duplex DNA.
Mechanism of Action of DNA Gyrase.
Mechanism of Action of Topoisomerase I.
Formation of Catenated DNAs.
Enzymatic Methods to Measure Topoisomerase I Activity.
Inserting DNA into a Cloning Vector.
Restriction Endonucleases.
DNA Ligases.
Phosphatase Treatment of Cloning Vectors.
Inter- and Intramolecular Joining of DNA Ends.
Cloning Strategies.
Insertional Inactivation.
Directional Cloning.
Replacement Cloning.
Construction of Recombinant DNA Libraries.
Lambda Libraries.
Lambda as a Cloning Vector.
Lambda ZAP II, an Insertion Vector.
cDNA Synthesis.
Replacement Vectors.
Purposes for Genomic and cDNA Cloning.
Methods for Detection of Clones.
Hybridization Methods.
Plaque Hybridization.
Colony Hybridization.
Histochemical Detection of Beta-Galactosidase.
Alpha Complementation.
pBluescript II SK-.
Antibody Screening Methods.
The BCIP/NBT Color Development Reaction for Antibody Detection.
M13 as a Cloning Vector.
Cloning Vectors of M13 and the Role of the lac Operon.
Comparison of M13 and Lambda.
The Life Cycle of M13 and Its Mode of Replication.
Phagemids.
DNA Sequencing and in Vitro Mutagenesis.
Enzymatic and Chemical Methods of DNA Sequence Analysis.
The Enzymatic Method.
The Chemical Method.
Laboratory Experiments.
Appendices.
Subject Index.