An Introduction to Molecular Biotechnology

Die Neuauflage dieses überaus renommierten Lehrbuchs wurde als Antwort auf die rasanten Fortschritte in dem Fachgebiet vollständig aktualisiert und präsentiert neue leistungsstarke Methoden und Konzepte in der Biotechnologie, u.a. Genome Editing, reprogrammierte Stammzellen und personalisierte Medizin.
Auf eine Einführung in die Grundlagen der Molekular- und Zellbiologie folgt eine Beschreibung der Standardverfahren, darunter Aufreinigung und Analyse von Biomolekülen, Verfahren der Klonierung, Gen-Expressionssysteme, Methoden des Genome Editing, Protein-Labeling und In-situ-Verfahren, Standard- und hochauflösende Mikroskopie. Der dritte Teil legt den Schwerpunkt auf wichtige Forschungs- und Anwendungsgebiete, von der funktionalen Genomik, Proteomik und Bioinformatik bis hin zu Drug Targeting, rekombinante Antikörper und Systembiologie. Der letzte Teil wirft einen Blick auf Unternehmen der Biotechnologie und untersucht Fragestellungen des geistigen Eigentums, den Rechtsrahmen für pharmazeutische Produkte und das Zusammenspiel von Startup- und größeren Unternehmen. Die Inhalte sind durchgängig überaus ansprechend illustriert, mit Hunderten von farbigen Diagrammen und Fotos.
Dieses Lehrbuch vermittelt Studenten und Berufspraktikern der Biowissenschaften, Pharmazie und Biochemie alles Wissenswerte rund um die molekulare Biotechnologie.

Auteur
Michael Wink studied biology and chemistry in Bonn and was awarded his doctorate from TU Braunschweig in 1980. After gaining his lecturing qualification in 1984/1985, he was awarded a Heisenberg grant by the German Research Council to work at the Max Planck Institute for Breeding Research in Cologne and from then at the Gene Center of Ludwig-Maximilians University in Munich. Following a chair for Pharmaceutical Biology at Mainz University in 1988, he accepted the post of Professor for Pharmaceutical Biology at the University of Heidelberg one year later. His areas of interest include pharmaceutical research, molecular biotechnology, and medicinal plants, as well as research into natural products and evolution.

Résumé
Completely updated in line with the rapid progress made in the field, this new edition of the highly-praised textbook addresses powerful new methods and concepts in biotechnology, such as genome editing, reprogrammed stem cells, and personalized medicine.
An introduction to the fundamentals in molecular and cell biology is followed by a description of standard techniques, including purification and analysis of biomolecules, cloning techniques, gene expression systems, genome editing methods, labeling of proteins and in situ-techniques, standard and high resolution microscopy. The third part focuses on key areas in research and application, ranging from functional genomics, proteomics and bioinformatics to drug targeting, recombinant antibodies and systems biology. The final part looks at the biotechnology industry, explaining intellectual property issues, legal frameworks for pharmaceutical products and the interplay between start-up and larger companies. The contents are beautifully illustrated throughout, with hundreds of full color diagrams and photographs.
Provides students and professionals in life sciences, pharmacy and biochemistry with everything they need to know about molecular biotechnology.

Contenu

Abbreviations xix

Part I Fundamentals of Cellular and Molecular Biology 1

1 The Cell as the Basic Unit of Life **3
**Michael Wink

References 8

Further Reading 8

**2 Structure and Function of Cellular Macromolecules 9
**Michael Wink

2.1 Structure and Function of Sugars 9

2.2 Structure of Membrane Lipids 13

2.3 Structure and Function of Proteins 17

2.4 Structure of Nucleotides and Nucleic Acids (DNA and RNA) 25

References 32

Further Reading 32

3 Structure and Functions of a Cell **33
**Michael Wink

3.1 Structure of a Eukaryotic Cell 33

3.1.1 Structure and Function of the Cytoplasmic Membrane 33

3.1.1.1 Membrane Permeability 33

3.1.1.2 Transport Processes Across Biomembranes 34

3.1.1.3 Receptors and Signal Transduction at Biomembranes 37

3.1.2 Endomembrane System in a Eukaryotic Cell 40

3.1.3 Mitochondria and Chloroplasts 45

3.1.4 Cytoplasm 49

3.1.5 Cytoskeleton 51

3.1.6 Cell Walls 53

3.2 Structure of Bacteria 53

3.3 Structure of Viruses 55

3.4 Differentiation of Cells 56

3.5 Cell Death 60

References 61

Further Reading 61

4 Biosynthesis and Function of Macromolecules (DNA, RNA, and Proteins) **63
**Michael Wink

4.1 Genomes, Chromosomes, and Replication 63

4.1.1 Genome Size 63

4.1.2 Composition and Function of Chromosomes 67

4.1.3 Mitosis and Meiosis 69

4.1.4 Replication 71

4.1.5 Mutations and Repair Mechanisms 72

4.2 Transcription: From Gene to Protein 77

4.3 Protein Biosynthesis (Translation) 81

Further Reading 85

5 Distributing Proteins in the Cell (Protein Sorting) **87
**Michael Wink

5.1 Import and Export of Proteins via the Nuclear Pore 87

5.2 Import of Proteins in Mitochondria, Chloroplasts, and Peroxisomes 88

5.3 Protein Transport into the Endoplasmic Reticulum 89

5.4 Vesicle Transport from the ER via the Golgi Apparatus to the Cytoplasmic Membrane 92

References 94

Further Reading 94

6 Evolution and Diversity of Organisms **95
**Michael Wink

6.1 Prokaryotes 95

6.2 Eukaryotes 95

References 101

Further Reading 101

Part II Standard Methods in Molecular Biotechnology 103

7 Isolation and Purification of Proteins **105
**Thomas Wieland

7.1 Introduction 105

7.2 Producing a Protein Extract 106

7.3 Gel Electrophoretic Separation Methods 107

7.3.1 Principles of Electrophoresis 107

7.3.2 Native Gel Electrophoresis 107

7.3.3 Discontinuous Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) 107

7.3.4 Two-Dimensional (2D) Gel Electrophoresis and Isoelectric Focusing (IEF) 108

7.3.5 Detecting Proteins in Gels 108

7.4 Methods of Protein Precipitation 109

7.5 Column Chromatography Methods 109

7.5.1 General Principles of Separation 109

7.5.1.1 Size Exclusion Chromatography (Gel Filtration) 109

7.5.1.2 Hydrophobic Interaction Chromatography 111

7.5.1.3 Ion Exchange Chromatography 111

7.5.1.4 Hydroxyapatite Chromatography 112

7.5.2 Group-Specific Separation Techniques 112

7.5.2.1 Chromatography on Protein A or Protein G 112

7.5.2.2 Chromatography on Cibacron Blue (Blue Gel) 112

7.5.2.3 Chromatography on Lectins 112

7.5.2.4 Chromatography on Heparin 113

7.5.3 Purification of Recombinant Fusion Proteins 113 7.5.3.1 Chromat...