Title Details

Preface XIX

List of Contributors XXI

Abbreviations XXV

Part I Fundamentals of Cellular and Molecular Biology 1

1 The Cell as the Basic Unit of Life 3
M. Wink

2 Structure and Function of Cellular Macromolecules 7
M. Wink

2.1 Structure and Function of Sugars 8

2.2 Structure of Membrane Lipids 10

2.3 Structure and Function of Proteins 14

2.4 Structure of Nucleotides and Nucleic Acids (DNA and RNA) 21

2.5 References 27

3 Structure and Functions of a Cell 29
M. Wink

3.1 Structure of a Eukaryotic Cell 29

3.2 Structure of Bacteria 50

3.3 Structure of Viruses 51

3.4 Differentiation of Cells 52

4 Biosynthesis and Function of Macromolecules (DNA, RNA, and Proteins) 57
M. Wink

4.1 Genomes, Chromosomes, and Replication 57

4.2 Transcription: From Gene to Protein 71

4.3 Protein Biosynthesis (Translation) 76

5 Distributing Proteins in the Cell (Protein Sorting) 81
M. Wink

5.1 Import and Export of Proteins via the Nuclear Pore 82

5.2 Import of Proteins in Mitochondria and Chloroplasts 83

5.3 Protein Transport into the Endoplasmic Reticulum 85

5.4 Vesicle Transport from the ER via the Golgi Apparatus to the Cytoplasmic Membrane 86

6 Evolution and Diversity of Organisms 91
M. Wink

6.1 Prokaryotes 91

6.2 Eukaryotes 91

Part II Standard Methods in Molecular Biotechnology 99

7 Isolation and Purification of Proteins 101
T. Wieland, M. Lutz

7.1 Introduction 101

7.2 Producing a Protein Extract 102

7.3 Gel Electrophoretic Separation Methods 103

7.4 Methods of Protein Precipitation 106

7.5 Column Chromatography Methods 107

7.6 Examples 113

8 Peptide and Protein Analysis with Electrospray Tandem Mass Spectrometry 115
A. Schlosser, W.D. Lehmann

8.1 Introduction 115

8.2 Principles of Mass Spectrometry 115

8.3 Mass Precision, Resolution, and Isotope Distribution 116

8.4 Principles of ESI 116

8.5 Tandem Mass Spectrometers 117

8.6 Peptide Sequencing with MS/MS 119

8.7 Identifying Proteins with MS/MS Data and Protein Databases 120

8.8 Determining Protein Molecular Mass 121

8.9 Analysis of Covalent Protein Modification 122

8.10 Relative and Absolute Quantification 123

9 Isolation of DNA and RNA 125
H. Weiher, R. Zwacka, I. Herr

9.1 Introduction 125

9.2 DNA Isolation 125

9.3 RNA Isolation 127

10 Chromatography and Electrophoresis of Nucleic Acids 129
H. Weiher, R. Zwacka, I. Herr

10.1 Introduction 129

10.2 Chromatographic Separation of Nucleic Acids 129

10.3 Electrophoresis 130

11 Hybridization of Nucleic Acids 133
H. Weiher, R. Zwacka, I. Herr

11.1 Significance of Base Pairing 133

11.2 Experimental Hybridization: Kinetic and Thermodynamic Control 133

11.3 Analytical Techniques 134

12 Use of Enzymes in the Modification of Nucleic Acids 137
A. Groth, R. Zwacka, H. Weiher, I. Herr

12.1 Restriction Enzymes (Restriction Endonucleases) 137

12.2 Ligases 139

12.3 Methyltransferases 139

12.4 DNA Polymerases 140

12.5 RNA Polymerases and Reverse Transcriptase 141

12.6 Nucleases 141

12.7 T4 Polynucleotide Kinase 141

12.8 Phosphatases 142

13 Polymerase Chain Reaction 143
A. Mohr, H. Weiher, I. Herr, R. Zwacka

13.1 Introduction 143

13.2 Techniques 143

13.3 Areas of Application 146

14 DNA Sequencing 149
R. Zwacka, A. Mohr, I. Herr, H. Weiher

14.1 Introduction 149

14.2 DNA Sequencing Methods 149

14.3 Strategies for Sequencing the Human Genome 151

14.4 Practical Significance of DNA 152

15 Cloning Procedures 153
T. Wieland, S. Lutz

15.1 Introduction 153

15.2 Construction of Recombinant Vectors 153

16 Expression of Recombinant Proteins 169
T. Wieland, S. Lutz

16.1 Introduction 169

16.2 Expression of Recombinant Proteins in Host Organisms 170

16.3 Expression in Cell-Free Systems 178

17 Patch Clamp Method 181
R. Kraft

17.1 Biological Membranes and Ion Channels 181

17.2 Physical Foundations of the Patch Clamp Method 182

17.3 Patch Clamp Configurations 182

17.4 Applications of the Patch Clamp Method 184

18 Cell Cycle Analysis 187
S. Wölfl, A. Kitanovic

18.1 Analyzing the Cell Cycle 187

18.2 Experimental Analysis of the Cell Cycle 189

19 Microscopic Techniques 197
S. Diekmann

19.1 Electron Microscopy 197

19.2 Atomic or Scanning Force Microscopy 200

19.3 Light Microscopy 202

19.4 Microscopy in the Living Cell 206

20 Laser Applications 211
M. Vogel, R. Fink

20.1 Principles of Laser Technology 211

20.2 Properties of Laser Radiation 213

20.3 Types of Lasers and Setups 213

20.4 Applications 214

Part III Key Topics 217

21 Genomics and Functional Genomics 219
S.Wiemann, M. Frohme

21.1 Introduction 219

21.2 Technological Developments in DNA Sequencing 221

21.3 Genome Sequencing 222

21.4 cDNA Projects 238

21.5 Functional Genomics 246

21.6 Identification and Analysis of Individual Genes 248

21.7 Investigation of Transcriptional Activity 253

21.8 Cell-based Methods 266

21.9 Functional Analysis of Entire Genomes 272

22 Bioinformatics 275
B. Brors, K. Fellenberg

22.1 Introduction 275

22.2 Data Sources 276

22.3 Sequence Analysis 279

22.4 Evolutionary Bioinformatics 283

22.5 Gene Prediction 287

22.6 Bioinformatics in Transcriptome and Proteome Analysis 288

22.7 Bioinformatic Software 293

23 Cellular Systems Biology 295
H. Schmidt-Gienewinkel, S. Legewie, B. Brors, R. König

23.1 Introduction 295

23.2 Analysis of Cellular Networks by Top-Down Approaches 296

23.3 Overview of Bottom-Up Modeling of Biochemical Networks 304

23.4 Biological Examples 309

24 Protein–Protein and Protein–DNA Interaction 315
P. Uetz, E. Pohl

24.1 Protein–Protein Interactions 315

24.2 Protein–DNA Interactions 324

25 Drug Research 331
M. Koegl, R. Tolle, U. Deuschle, C. Kremoser

25.1 Introduction 331

25.2 Active Compounds and their Targets 331

25.3 Preclinical Pharmacology and Toxicology 344

25.4 Clinical Development 346

25.5 Clinical Testing 346

26 Drug Targeting and Prodrugs 349
G. Fricker

26.1 Drug Targeting 349

26.2 Prodrugs 355

26.3 Penetration of Drugs through Biological Membranes 356

26.4 Prodrugs to Extend Duration of Effect 357

26.5 Prodrugs for the Targeted Release of a Drug 357

26.6 Prodrugs to Minimize Side Effects 358

27 Molecular Diagnostics in Medicine 359
S. Wölfl, R. Gessner

27.1 Uses of Molecular Diagnostics 359

27.2 Which Molecular Variations Should be Detected 364

27.3 Molecular Diagnostic Methods 367

27.4 Outlook 375

28 Recombinant Antibodies and Phage Display 377
S. Dübel

28.1 Introduction 377

28.2 Why Recombinant Antibodies? 379

28.2.1 Recombinant Antibodies are Available In Vitro without Immunization 379

28.3 Obtaining Specific Recombinant Antibodies 379

28.4 Production of Recombinant Antibodies 384

28.5 Formats for Recombinant Antibodies 386

28.6 Applications of Recombinant Antibodies 392

28.7 Outlook 394

29 Transgenic and Gene-Targeted Mice and their Impact in Medical Research 395
R. Sprengel

29.1 Overview 395

29.2 Transgenic Mice 395

29.3 Homologous Recombination: knock-out (-in) mice 398

29.4 Conditionally Regulated Gene Expression 399

29.5 Impact of Genetically Modified Mice in Biomedicine 400

29.6 Outlook 402

30 Gene Therapy: Strategies and Vectors 403
A. Groth, I. Herr

30.1 Introduction 403

30.2 Principles of Somatic Gene Therapy 404

30.3 Germ Line Therapy 405

30.4 Setbacks in Gene Therapy 406

30.5 Vectors for Gene Therapy 406

30.6 Specific Expression 413

31 RNA Interference, Modified DNA, Peptide Nucleic Acid, and Applications in Medicine and Biotechnology 415
N. Metzler-Nolte, A. Sosniak

31.1 Introduction 415

31.2 Modified Nucleic Acids 416

31.3 Interactions of DNA Analogs with Complementary DNA and RNA 419

31.4 RNAi 421

31.5 Applications 424

32 Plant Biotechnology 431
H. Hillebrand, R. Hell

32.1 Introduction 431

32.2 Gene Expression Control 433

32.3 Production of Transgenic Plants 434

32.4 Selection of Transformed Plant Cells 439

32.5 Regeneration of Transgenic Plants 445

32.6 Plant Analysis: Identification and Characterization of Genetically Engineered Plants 446

33 Biocatalysis in the Chemical Industry 451
M. Breuer, B. Hauer

33.1 Introduction 451

33.2 Bioconversion/Enzymatic Procedures 454

33.3 Development of an Enzyme for Industrial Biocatalysis 456

33.4 Fermentative Procedures 462

Part IV Biotechnology in Industry 473

34 Industrial Application: Biotech Industry, Markets, and Opportunities 475
J. Schüler

34.1 Historical Overview and Definitions of Concepts 475

34.2 Areas of Industrial Application of Molecular Biotechnology 476

34.3 Status Quo of the Biotech Industry World-Wide 485

35 Patents in the Molecular Biotechnology Industry: Legal and Ethical Issues 487
David B. Resnik

35.1 Patent Law 487

35.2 Ethical and Policy Issues in Biotechnology Patents 492

35.3 Conclusions 498

36 Drug Approval in the European Union and United States 499
G. Walsh

36.1 Introduction 499

36.2 Regulation within the European Union 499

36.3 Regulation in the United States 503

36.4 Advent and Regulation of Biosimilars 506

36.5 International Regulatory Harmonization 506

37 Emergence of a Biotechnology Industry 509
C. Kremoser

38 The 101 of Founding a Biotech Company 517
C. Kremoser

38.1 First Steps Towards Your Own Company 517

38.2 Employees: Recruitment, Remuneration, Participation 522

39 Marketing 527
C. Kremoser

39.1 Introduction 527

39.2 What Types of Deals are Possible? 528

39.3 What Milestone or License Fees are Effectively Paid in a Biotech/Pharma Cooperation? 529

39.4 PR and IR in Biotech Companies 530

Appendix 533

Further Reading 535

Glossary 551
M. Wink

Subject Index 587

"Overall, however, I am very satisfied with the contents of this book (I wish I could edit such a nice book !), and would like to recommend it to the undergraduate and graduate students alike who want to quickly yet thoroughly learn the fundamentals and many applications of molecular biotechnology. Also, this book will be a nice and smooth reading for professors, researchers, and professionals in the field of bioscience and biotechnology to update themselves who wish to catch up with the recent developments in molecular biotechnology. In conclusion, this book will be a must read for modern bioscientists and biotechnologists and those who want to become." (Chemical and Biomolecular Engineering, 1 May 2011)

"This book is a must read for modern bioscientists and biotechnologists and those who want to become one.” (Biotechnology Journal, 1 July 2011)