Principles and techniques of practical biochemistry fifth editionPDF电子书下载

1 General principles of biochemical investigations&I.SIMPKINS 1

1.1 The nature of biochemistry 1

1.2 Bioenergetics 4

1.3 Methods for investigating metabolism 21

1.4 Practical considerations 28

1.5 In vivomodels 42

1.6 Invitromodels 44

1.7 Microscopy 66

1.8 Key terms 75

1.9 Calculations 76

1.10 Suggestions for further reading 78

2 Molecular biology and basic techniques&R.RAPLEY 80

2.1 Introduction 80

2.2 Components and primary structure of nucleic acids 80

2.3 Genes and genome complexity 87

2.4 The nature of the genetic code 90

2.5 Cellular location of nucleic acids 90

2.6 The cellular functions of DNA 93

2.7 The manipulation of nucleic acids：basic tools and techniques 103

2.8 Isolation and separation of nucleic acids 105

2.9 Restriction mapping of DNA fragments 110

2.10 Nucleic acid blotting methods 111

2.11 Gene probe derivation 113

2.12 Labelling DNA gene probe molecules 114

2.13 The polymerase chain reaction 116

2.14 Nucleotide sequencing of DNA 125

2.15 Bioinformatics and the Internet 131

2.16 Key terms 134

2.17 Calculations 135

2.18 Suggestions for further reading 136

3 Molecular cloning and gene analysis&R.RAPLEY 138

3.1 Introduction 138

3.2 Constructing gene libraries 138

3.3 Cloning vectors 148

3.4 Hybridisation and gene probes 167

3.5 Screening gene libraries 167

3.6 Applications of gene cloning 171

3.7 Expression of foreign genes 178

3.8 Analysing genes and gene expression 184

3.9 Analysing whole genomes 196

3.10 Molecular biotechnology and applications 202

3.11 Key terms 204

3.12 Suggestions for further reading 205

4 Immunochemical techniques&R.THORPE AND S.THORPE 206

4.1 Introduction 206

4.2 Production of antibodies 211

4.3 Purification and fragmentation of immunoglobulins 222

4.4 Immunoprecipitation 229

4.5 Labelling antibodies 234

4.6 Immunoblotting 240

4.7 Immunoassays 244

4.8 Immunohisto/cytochemistry 254

4.9 Affinity and avidity 260

4.10 Immunochemical use of surface plasmon resonance 260

4.11 Key terms 261

4.12 Calculation 262

4.13 Suggestions for further reading 262

5 Centrifugation techniques&A.GRIFFITHS 263

5.1 Introduction 263

5.2 Basic principles of sedimentation 264

5.3 Centrifuges and their uses 271

5.4 Design and care of preparative rotors 276

5.5 Sample containers 284

5.6 Separation methods in preparative ultracentrifuges 285

5.7 Performing density gradient separations 290

5.8 Selection，efficiency and applications of preparative rotors 296

5.9 Analysis of subcellular fractions 300

5.10 Some applications of the analytical ultracentrifuge 302

5.11 Safety aspects in the use of centrifuges 305

5.12 Key terms 306

5.13 Calculations 306

5.14 Suggestions for further reading 311

6 Protein structure，purification and characterisation&J.WALKER 312

6.1 Ionic properties of amino acids and proteins 312

6.2 Protein structure 316

6.3 Protein purification 318

6.4 Protein structure determination 338

6.5 Key terms 353

6.6 Calculations 354

6.7 Suggestions for further reading 356

7 Biomolecular interactions：Ⅰ Enzymes&K.WILSON 357

7.1 Receptor-ligand binding 357

7.2 Enzymes：characteristics and nomenclature 358

7.3 Enzyme steady-state kinetics 359

7.4 Enzyme assays 378

7.5 Substrate assays 385

7.6 Enzyme pre-steady-state kinetics 386

7.7 Enzyme active sites and catalytic mechanisms 389

7.8 Immobilised enzymes 394

7.9 Cellular control of metabolic activity 396

7.10 Key terms 400

7.11 Calculations 401

7.12 Suggestions for further reading 402

8 Biomolecular interactions：Ⅱ Cell surface receptors and transporters&K.WILSON 403

8.1 Cell surface receptor classification 403

8.2 Quantitative aspects of receptor-ligand binding 406

8.3 Receptor structures 417

8.4 Mechanisms of signal transduction 420

8.5 Signal amplification 430

8.6 Key terms 432

8.7 Membrane transport processes 432

8.8 Physical diffusion 433

8.9 Facilitated transport 435

8.10 Active transport and ion channels 438

8.11 Receptor-mediated endocytosis 445

8.12 Key terms 449

8.13 Calculations 449

8.14 Suggestions for further reading 451

9 Spectroscopic techniques：Ⅰ Atomic and molecular electronic spectroscopy&D.B.GORDON 453

9.1 Introduction 453

9.2 γ-Ray spectroscopy and γ-ray resonance spectroscopy 456

9.3 X-ray spectroscopy 458

9.4 Ultraviolet and visible light spectroscopy 459

9.5 Spectrofluorimetry 471

9.6 Circular dichroism spectroscopy 479

9.7 Turbidimetry and nephelometry 482

9.8 Luminometry 483

9.9 Atomic spectroscopy 485

9.10 Lasers 490

9.11 Key terms 491

9.12 Calculations 492

9.13 Suggestions for further reading 497

10 Spectroscopic techniques：Ⅱ Vibrational spectroscopy and electron and nuclear spin orientation in magnetic fieldsz&D.B.GORDON 498

10.1 Introduction 498

10.2 Infrared and Raman spectroscopy 498

10.3 Electron spin resonance spectroscopy 501

10.4 Nuclear magnetic resonance spectroscopy 508

10.5 Key terms 525

10.6 Suggestions for further reading 526

11 Mass spectrometric techniques&D.B.GORDON 527

11.1 Introduction 527

11.2 The mass spectrometer 527

11.3 Electron impact ionisation 529

11.4 Chemical ionisation 538

11.5 Field ionisation 539

11.6 Ion desorption methods 539

11.7 Ion evaporation methods 551

11.8 Analysers 555

11.9 Detectors 564

11.10 Tandem mass spectrometry 566

11.11 Key terms 573

11.12 Calculations 573

11.13 Suggestions for further reading 579

12 Electrophoretic techniques&J.M.WALKER 580

12.1 General principles 580

12.2 Support media 584

12.3 Electrophoresis of proteins 588

12.4 Electrophoresis ofnucleic acids 607

12.5 Capillary electrophoresis 612

12.6 Key terms 617

12.7 Calculation 618

12.8 Suggestions for further reading 618

13 Chromatographic techniques&K.WILSON 619

13.1 Introduction 619

13.2 Chromatography theory and practice 623

13.3 Low pressure column chromatography 631

13.4 High performance liquid chromatography 637

13.5 Adsorption chromatography 647

13.6 Partition chromatography 649

13.7 Ion-exchange chromatography 656

13.8 Molecular exclusion（permeation）chromatography 661

13.9 Affinity chromatography 665

13.10 Gas-liquid chromatography 672

13.11 Thin-layer（planar）chromatography 678

13.12 Selection of a chromatographic system 681

13.13 Key terms 682

13.14 Calculations 683

13.15 Suggestions for further reading 686

14 Radioisotope techniques&R.J.SLATER 687

14.1 The nature of radioactivity 687

14.2 Detection and measurement of radioactivity 693

14.3 Other practical aspects of counting radioactivity and analysis of data 713

14.4 Inherent advantages and restrictions of radiotracer experiments 717

14.5 Safety aspects 718

14.6 Applications of radioisotopes in the biological sciences 721

14.7 Key terms 726

14.8 Calculations 726

14.9 Suggestions for further reading 728

15 Electrochemical techniques&P.K.ROBINSON 729

15.1 Introduction 729

15.2 Principles of electrochemical techniques 734

15.3 Redox reactions 742

15.4 The pH electrode 745

15.5 Ion-selective and gas-sensing electrodes 748

15.6 The Clark oxygen electrode 750

15.7 Electrochemical detectors for HPLC 757

15.8 Biosensors 760

15.9 Key terms 768

15.10 Calculations 768

15.11 Suggestions for further reading 770

Index 771