<p>Preface XIX</p> <p>List of Contributors XXI</p> <p>Abbreviations XXV</p> <p><b>Part I Fundamentals of Cellular and Molecular Biology 1</b></p> <p><b>1 The Cell as the Basic Unit of Life 3</b><br /><i>M. Wink</i></p> <p><b>2 Structure and Function of Cellular Macromolecules 7</b><br /><i>M. Wink</i></p> <p>2.1 Structure and Function of Sugars 8</p> <p>2.2 Structure of Membrane Lipids 10</p> <p>2.3 Structure and Function of Proteins 14</p> <p>2.4 Structure of Nucleotides and Nucleic Acids (DNA and RNA) 21</p> <p>2.5 References 27</p> <p><b>3 Structure and Functions of a Cell 29</b><br /><i>M. Wink</i></p> <p>3.1 Structure of a Eukaryotic Cell 29</p> <p>3.2 Structure of Bacteria 50</p> <p>3.3 Structure of Viruses 51</p> <p>3.4 Differentiation of Cells 52</p> <p><b>4 Biosynthesis and Function of Macromolecules (DNA, RNA, and Proteins) 57</b><br /><i>M. Wink</i></p> <p>4.1 Genomes, Chromosomes, and Replication 57</p> <p>4.2 Transcription: From Gene to Protein 71</p> <p>4.3 Protein Biosynthesis (Translation) 76</p> <p><b>5 Distributing Proteins in the Cell (Protein Sorting) 81</b><br /><i>M. Wink</i></p> <p>5.1 Import and Export of Proteins via the Nuclear Pore 82</p> <p>5.2 Import of Proteins in Mitochondria and Chloroplasts 83</p> <p>5.3 Protein Transport into the Endoplasmic Reticulum 85</p> <p>5.4 Vesicle Transport from the ER via the Golgi Apparatus to the Cytoplasmic Membrane 86</p> <p><b>6 Evolution and Diversity of Organisms 91</b><br /><i>M. Wink</i></p> <p>6.1 Prokaryotes 91</p> <p>6.2 Eukaryotes 91</p> <p><b>Part II Standard Methods in Molecular Biotechnology 99</b></p> <p><b>7 Isolation and Purification of Proteins 101</b><br /><i>T. Wieland, M. Lutz</i></p> <p>7.1 Introduction 101</p> <p>7.2 Producing a Protein Extract 102</p> <p>7.3 Gel Electrophoretic Separation Methods 103</p> <p>7.4 Methods of Protein Precipitation 106</p> <p>7.5 Column Chromatography Methods 107</p> <p>7.6 Examples 113</p> <p><b>8 Peptide and Protein Analysis with Electrospray Tandem Mass Spectrometry 115</b><br /><i>A. Schlosser, W.D. Lehmann</i></p> <p>8.1 Introduction 115</p> <p>8.2 Principles of Mass Spectrometry 115</p> <p>8.3 Mass Precision, Resolution, and Isotope Distribution 116</p> <p>8.4 Principles of ESI 116</p> <p>8.5 Tandem Mass Spectrometers 117</p> <p>8.6 Peptide Sequencing with MS/MS 119</p> <p>8.7 Identifying Proteins with MS/MS Data and Protein Databases 120</p> <p>8.8 Determining Protein Molecular Mass 121</p> <p>8.9 Analysis of Covalent Protein Modification 122</p> <p>8.10 Relative and Absolute Quantification 123</p> <p><b>9 Isolation of DNA and RNA 125</b><br /><i>H. Weiher, R. Zwacka, I. Herr</i></p> <p>9.1 Introduction 125</p> <p>9.2 DNA Isolation 125</p> <p>9.3 RNA Isolation 127</p> <p><b>10 Chromatography and Electrophoresis of Nucleic Acids 129</b><br /><i>H. Weiher, R. Zwacka, I. Herr</i></p> <p>10.1 Introduction 129</p> <p>10.2 Chromatographic Separation of Nucleic Acids 129</p> <p>10.3 Electrophoresis 130</p> <p><b>11 Hybridization of Nucleic Acids 133</b><br /><i>H. Weiher, R. Zwacka, I. Herr</i></p> <p>11.1 Significance of Base Pairing 133</p> <p>11.2 Experimental Hybridization: Kinetic and Thermodynamic Control 133</p> <p>11.3 Analytical Techniques 134</p> <p><b>12 Use of Enzymes in the Modification of Nucleic Acids 137</b><br /><i>A. Groth, R. Zwacka, H. Weiher, I. Herr</i></p> <p>12.1 Restriction Enzymes (Restriction Endonucleases) 137</p> <p>12.2 Ligases 139</p> <p>12.3 Methyltransferases 139</p> <p>12.4 DNA Polymerases 140</p> <p>12.5 RNA Polymerases and Reverse Transcriptase 141</p> <p>12.6 Nucleases 141</p> <p>12.7 T4 Polynucleotide Kinase 141</p> <p>12.8 Phosphatases 142</p> <p><b>13 Polymerase Chain Reaction 143</b><br /><i>A. Mohr, H. Weiher, I. Herr, R. Zwacka</i></p> <p>13.1 Introduction 143</p> <p>13.2 Techniques 143</p> <p>13.3 Areas of Application 146</p> <p><b>14 DNA Sequencing 149</b><br /><i>R. Zwacka, A. Mohr, I. Herr, H. Weiher</i></p> <p>14.1 Introduction 149</p> <p>14.2 DNA Sequencing Methods 149</p> <p>14.3 Strategies for Sequencing the Human Genome 151</p> <p>14.4 Practical Significance of DNA 152</p> <p><b>15 Cloning Procedures 153</b><br /><i>T. Wieland, S. Lutz</i></p> <p>15.1 Introduction 153</p> <p>15.2 Construction of Recombinant Vectors 153</p> <p><b>16 Expression of Recombinant Proteins 169</b><br /><i>T. Wieland, S. Lutz</i></p> <p>16.1 Introduction 169</p> <p>16.2 Expression of Recombinant Proteins in Host Organisms 170</p> <p>16.3 Expression in Cell-Free Systems 178</p> <p><b>17 Patch Clamp Method 181</b><br /><i>R. Kraft</i></p> <p>17.1 Biological Membranes and Ion Channels 181</p> <p>17.2 Physical Foundations of the Patch Clamp Method 182</p> <p>17.3 Patch Clamp Configurations 182</p> <p>17.4 Applications of the Patch Clamp Method 184</p> <p><b>18 Cell Cycle Analysis 187</b><br /><i>S. Wölfl, A. Kitanovic</i></p> <p>18.1 Analyzing the Cell Cycle 187</p> <p>18.2 Experimental Analysis of the Cell Cycle 189</p> <p><b>19 Microscopic Techniques 197</b><br /><i>S. Diekmann</i></p> <p>19.1 Electron Microscopy 197</p> <p>19.2 Atomic or Scanning Force Microscopy 200</p> <p>19.3 Light Microscopy 202</p> <p>19.4 Microscopy in the Living Cell 206</p> <p><b>20 Laser Applications 211</b><br /><i>M. Vogel, R. Fink</i></p> <p>20.1 Principles of Laser Technology 211</p> <p>20.2 Properties of Laser Radiation 213</p> <p>20.3 Types of Lasers and Setups 213</p> <p>20.4 Applications 214</p> <p><b>Part III Key Topics 217</b></p> <p><b>21 Genomics and Functional Genomics 219</b><br /><i>S.Wiemann, M. Frohme</i></p> <p>21.1 Introduction 219</p> <p>21.2 Technological Developments in DNA Sequencing 221</p> <p>21.3 Genome Sequencing 222</p> <p>21.4 cDNA Projects 238</p> <p>21.5 Functional Genomics 246</p> <p>21.6 Identification and Analysis of Individual Genes 248</p> <p>21.7 Investigation of Transcriptional Activity 253</p> <p>21.8 Cell-based Methods 266</p> <p>21.9 Functional Analysis of Entire Genomes 272</p> <p><b>22 Bioinformatics 275</b><br /><i>B. Brors, K. Fellenberg</i></p> <p>22.1 Introduction 275</p> <p>22.2 Data Sources 276</p> <p>22.3 Sequence Analysis 279</p> <p>22.4 Evolutionary Bioinformatics 283</p> <p>22.5 Gene Prediction 287</p> <p>22.6 Bioinformatics in Transcriptome and Proteome Analysis 288</p> <p>22.7 Bioinformatic Software 293</p> <p><b>23 Cellular Systems Biology 295</b><br /><i>H. Schmidt-Gienewinkel, S. Legewie, B. Brors, R. König</i></p> <p>23.1 Introduction 295</p> <p>23.2 Analysis of Cellular Networks by Top-Down Approaches 296</p> <p>23.3 Overview of Bottom-Up Modeling of Biochemical Networks 304</p> <p>23.4 Biological Examples 309</p> <p><b>24 Protein–Protein and Protein–DNA Interaction 315</b><br /><i>P. Uetz, E. Pohl</i></p> <p>24.1 Protein–Protein Interactions 315</p> <p>24.2 Protein–DNA Interactions 324</p> <p><b>25 Drug Research 331</b><br /><i>M. Koegl, R. Tolle, U. Deuschle, C. Kremoser</i></p> <p>25.1 Introduction 331</p> <p>25.2 Active Compounds and their Targets 331</p> <p>25.3 Preclinical Pharmacology and Toxicology 344</p> <p>25.4 Clinical Development 346</p> <p>25.5 Clinical Testing 346</p> <p><b>26 Drug Targeting and Prodrugs 349</b><br /><i>G. Fricker</i></p> <p>26.1 Drug Targeting 349</p> <p>26.2 Prodrugs 355</p> <p>26.3 Penetration of Drugs through Biological Membranes 356</p> <p>26.4 Prodrugs to Extend Duration of Effect 357</p> <p>26.5 Prodrugs for the Targeted Release of a Drug 357</p> <p>26.6 Prodrugs to Minimize Side Effects 358</p> <p><b>27 Molecular Diagnostics in Medicine 359</b><br /><i>S. Wölfl, R. Gessner</i></p> <p>27.1 Uses of Molecular Diagnostics 359</p> <p>27.2 Which Molecular Variations Should be Detected 364</p> <p>27.3 Molecular Diagnostic Methods 367</p> <p>27.4 Outlook 375</p> <p><b>28 Recombinant Antibodies and Phage Display 377</b><br /><i>S. Dübel</i></p> <p>28.1 Introduction 377</p> <p>28.2 Why Recombinant Antibodies? 379</p> <p>28.2.1 Recombinant Antibodies are Available In Vitro without Immunization 379</p> <p>28.3 Obtaining Specific Recombinant Antibodies 379</p> <p>28.4 Production of Recombinant Antibodies 384</p> <p>28.5 Formats for Recombinant Antibodies 386</p> <p>28.6 Applications of Recombinant Antibodies 392</p> <p>28.7 Outlook 394</p> <p><b>29 Transgenic and Gene-Targeted Mice and their Impact in Medical Research 395</b><br /><i>R. Sprengel</i></p> <p>29.1 Overview 395</p> <p>29.2 Transgenic Mice 395</p> <p>29.3 Homologous Recombination: knock-out (-in) mice 398</p> <p>29.4 Conditionally Regulated Gene Expression 399</p> <p>29.5 Impact of Genetically Modified Mice in Biomedicine 400</p> <p>29.6 Outlook 402</p> <p><b>30 Gene Therapy: Strategies and Vectors 403</b><br /><i>A. Groth, I. Herr</i></p> <p>30.1 Introduction 403</p> <p>30.2 Principles of Somatic Gene Therapy 404</p> <p>30.3 Germ Line Therapy 405</p> <p>30.4 Setbacks in Gene Therapy 406</p> <p>30.5 Vectors for Gene Therapy 406</p> <p>30.6 Specific Expression 413</p> <p><b>31 RNA Interference, Modified DNA, Peptide Nucleic Acid, and Applications in Medicine and Biotechnology 415</b><br /><i>N. Metzler-Nolte, A. Sosniak</i></p> <p>31.1 Introduction 415</p> <p>31.2 Modified Nucleic Acids 416</p> <p>31.3 Interactions of DNA Analogs with Complementary DNA and RNA 419</p> <p>31.4 RNAi 421</p> <p>31.5 Applications 424</p> <p><b>32 Plant Biotechnology 431</b><br /><i>H. Hillebrand, R. Hell</i></p> <p>32.1 Introduction 431</p> <p>32.2 Gene Expression Control 433</p> <p>32.3 Production of Transgenic Plants 434</p> <p>32.4 Selection of Transformed Plant Cells 439</p> <p>32.5 Regeneration of Transgenic Plants 445</p> <p>32.6 Plant Analysis: Identification and Characterization of Genetically Engineered Plants 446</p> <p><b>33 Biocatalysis in the Chemical Industry 451</b><br /><i>M. Breuer, B. Hauer</i></p> <p>33.1 Introduction 451</p> <p>33.2 Bioconversion/Enzymatic Procedures 454</p> <p>33.3 Development of an Enzyme for Industrial Biocatalysis 456</p> <p>33.4 Fermentative Procedures 462</p> <p><b>Part IV Biotechnology in Industry 473</b></p> <p><b>34 Industrial Application: Biotech Industry, Markets, and Opportunities 475</b><br /><i>J. Schüler</i></p> <p>34.1 Historical Overview and Definitions of Concepts 475</p> <p>34.2 Areas of Industrial Application of Molecular Biotechnology 476</p> <p>34.3 Status Quo of the Biotech Industry World-Wide 485</p> <p><b>35 Patents in the Molecular Biotechnology Industry: Legal and Ethical Issues 487</b><br /><i>David B. Resnik</i></p> <p>35.1 Patent Law 487</p> <p>35.2 Ethical and Policy Issues in Biotechnology Patents 492</p> <p>35.3 Conclusions 498</p> <p><b>36 Drug Approval in the European Union and United States 499</b><br /><i>G. Walsh</i></p> <p>36.1 Introduction 499</p> <p>36.2 Regulation within the European Union 499</p> <p>36.3 Regulation in the United States 503</p> <p>36.4 Advent and Regulation of Biosimilars 506</p> <p>36.5 International Regulatory Harmonization 506</p> <p><b>37 Emergence of a Biotechnology Industry 509</b><br /><i>C. Kremoser</i></p> <p><b>38 The 101 of Founding a Biotech Company 517</b><br /><i>C. Kremoser</i></p> <p>38.1 First Steps Towards Your Own Company 517</p> <p>38.2 Employees: Recruitment, Remuneration, Participation 522</p> <p><b>39 Marketing 527</b><br /><i>C. Kremoser</i></p> <p>39.1 Introduction 527</p> <p>39.2 What Types of Deals are Possible? 528</p> <p>39.3 What Milestone or License Fees are Effectively Paid in a Biotech/Pharma Cooperation? 529</p> <p>39.4 PR and IR in Biotech Companies 530</p> <p>Appendix 533</p> <p>Further Reading 535</p> <p>Glossary 551<br />M. Wink</p> <p>Subject Index 587</p>