Brenda D. Spangler

Methods in Molecular Biology and Protein Chemistry

Cloning and Characterization of an Enterotoxin Subunit

• Broschiertes Buch

Produktbeschreibung

The enormous advances made in molecular biology have allowed scientists to manipulate DNA with relative ease. This means that molecular biology has become a widely-used tool for answering scientific questions that may be quite unrelated to genetics and cell biology.

This new book focuses on the most important techniques needed by undergraduate and post-graduate students when undertaking research in this area. Written in an accessible style and adopting a 'discovery' approach, to encourage the students to explore and experiment for them selves, this book will be invaluable to novices required and interested in learning these techniques.
_ Students will evaluate the effect of genetic engineering through the practical application of wet biochemistry techniques and prepare mutations that can be evaluated by other classes.
_ Coverage will include computer tutorials for sequencing and genomics and will possibly include some coverage of molecular modelling too.
_ Guidelines on searching the primary literature, oral reports, laboratory notebooks record keeping and report writing will either be included within appendices at the back of the book, or on a supplementary web site linked to the book.

Produktdetails

• Verlag: Wiley & Sons
• 1. Auflage
• Seitenzahl: 220
• Erscheinungstermin: 14. Juni 2002
• Englisch
• Abmessung: 280mm x 210mm x 12mm
• Gewicht: 642g
• ISBN-13: 9780470843604
• ISBN-10: 0470843608
• Artikelnr.: 10536330

Autorenporträt

Brenda Spangler has over 30 years experience in the field of protein chemistry and protein toxins, specializing in ligand-receptior molecular recognition.

Inhaltsangabe

Preface Introduction and Background Part 1 Molecular Biology Experiment 1
Instructions for Pipetman Use Experiment 2 Preparation of Plasmid Template
DNA Experiment 3 Estimation of Plasmid DNA Concentration PCR Amplification
of LTB Gene from Plasmid DNA Experiment 4 Agarose Gel Eletrophoresis
Recovery of PCR Product Experiment 5 Restriction Digest of LTB Insert Plate
Preparation Experiment 6 Purification of Digested Insert and Vector by
Agarose Gel Eletrophoresis Recovery of Digested Insert and Vector
Experiment 7 Determination of DNA Concentrations Ligation of Insert into
Vector Experiment 8 Transformation of Host Cells by Construct Experiment 9
Colony-pick PCR to Check for Presence of Insert Streak Plates with
Potential Constructs Inoculate Cultures/Run PCR Experiment 10 Agarose Gel
Electrophoresis to Verify Presence of Insert Purification of the Plasmid
Construct using a Plasmid Miniprep Kit Experiment 11 DNA Concentration
Determination Ethanol Precipitation of Plasmid DNA Sequencing Reaction
Experiment 12 Purification of Extension Reaction Products Sequencing Gel
Demonstration Experiment 13 Analysis of Sequencing Data Verification of
Insert Sequence Experiment 14 Gene Expression Transformation into
Expression Host Experiment 15 Induction of LTB Gene Expression Part 2
Protein Chemistry Experiment 16 SDS-PAGE of Induction Time Course Transfer
of Protein to Membrane for Western Blot Experiment 17 Visualization of
Western Blot Experiment 18 Large Scale Culture Preparation Experiment 19
Isolation of LTB by Affinity Chromatography Experiment 20 SDS-PAGE of
Column Fractions Immunoblot to verify Presence of LTB Experiment 21 Protein
Concentration Protein Crystallization Experiment 22 Setting up an ELISA
Microtiter Plate Experiment 23 ELISA of a Panel of Ganglioside Ligands
Experiment 24 Protein Structure Determination: X-ray Diffraction Techniques
Experiment 25 Characterization of a Protein Crystal Experiment 26 Primer
Design