实用生物化学的原理与技术(第5版.英文版)

.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 nucleotidesequencingofdna 125
2.15 bioinformatics and the internet 131
2.16 keyterms 134
2.17 calculations 135
2.18 suggestions for further reading 136
3 molecular cloning and gene analysis 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 molecularbiotechnologyandapplications 202
3.11 keyterms 204
3.12 suggestions for further reading 205
4 immunochemicaltechniques 206
4.1 introduction 206
4.2 production of antibodies 211
4.3 purification and fragmentation ofimmunoglobulins 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 k eyterms 261
4.12 calculation 262
4.13 suggestions for further reading 262
5 centrifugationtechniques 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, efficiencyandapplicationsofpreparativerotors 296
5.9 analysis of subcellular fractions 300
5.10 someapplicationsoftheanalyticalultracentrifuge 302
5.11 safety aspects in the use of centrifuges 305
5.12 keyterms 306
5.13 calculations 306
5.14 suggestions for further reading 311
6 protein structure, purification and characterisation 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 357
7.1 receptor-ligandbinding 357
7.2 enzymes: characteristics and nomenclature 358
7.3 enzyme steady-state kinetics 359
7.4 enzyme assays 3 78
7.5 substrate assays 385
7.6 enzyme pre-steady-state kinetics 386
7.7 enzyme active sites and catalytic mechanisms 389
7.8 immobilisedenzymes 394
7.9 cellular control ofmetabolic activity 396
7.10 keyterms 400
7.11 calculations .. 401
7.12 suggestions for further reading 402
8 8iomolecular interactions: ii cell surface receptors and transporters 403
8.1 cellsurface receptor classification 403
8.2 quantitative aspects ofreceptor-ligand binding 406
8.3 receptorstructures 417
8.4 mechanismsofsignaltransduction 420
8.5 signal amplification 430
8.6 key terms 432
8.7 membrane transport processes 432
8.8 physicaldiffusion 433
8.9 facilitated transport 435
8.10 active transport and ion channels 438
8.11 receptor-mediatedendocytosis 445
8.12 keyterms 449
8.13 calculations 449
8.14 suggestions for further reading 451
9 spectroscopic techniques: i atomic and molecular electronic spectroscopy 453
9.1 introduction 453
9.2 y-ray spectroscopy and y-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 keyterms 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 fields 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 527
11.1 introduction 527
11.2 the mass spectrometer 527
11.3 electron impact ionisation 529
11.4 chemical ionisation 538
11.5 fieldionisation 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 furtherreading 579
12 electrophoretic techniques 580
12.1 general principles 580
12.2 support media 584
12.3 electrophoresis of proteins 588
12.4 electrophoresis of nucleic 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 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 ofa chromatographic system 681
13.13 key terms 682
13.14 calculations 683
13.15 suggestions for further reading 686
14 radioisotope techniques 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 729
15.1 introduction 729
15.2 principles of electrochemicaltechniques 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