Gene cloning and DNA analysis: an introduction
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| Format: | Book |
| Language: | English |
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Hoboken, NJ ; Chichester
Wiley Blackwell
2021
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| Edition: | Eighth edition |
| Subjects: | |
| Online Access: | Inhaltsverzeichnis |
| Physical Description: | xv, 416 Seiten Illustrationen, Diagramme, Karten |
| ISBN: | 9781119640783 |
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| 100 | 1 | |a Brown, Terence A. |d 1953- |e Verfasser |0 (DE-588)113251661 |4 aut | |
| 245 | 1 | 0 | |a Gene cloning and DNA analysis |b an introduction |c T. A. Brown (University of Manchester, Manchester, United Kingdom) |
| 250 | |a Eighth edition | ||
| 264 | 1 | |a Hoboken, NJ ; Chichester |b Wiley Blackwell |c 2021 | |
| 300 | |a xv, 416 Seiten |b Illustrationen, Diagramme, Karten | ||
| 336 | |b txt |2 rdacontent | ||
| 337 | |b n |2 rdamedia | ||
| 338 | |b nc |2 rdacarrier | ||
| 650 | 2 | |a Cloning, Molecular | |
| 650 | 2 | |a DNA, Recombinant - analysis | |
| 650 | 2 | |a Sequence Analysis, DNA | |
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| 653 | |a Genomforschung u. Proteomik | ||
| 653 | |a Gentechnologie | ||
| 653 | |a Genomics & Proteomics | ||
| 653 | |a Genetik | ||
| 653 | |a Genetics | ||
| 653 | |a Cell & Molecular Biology | ||
| 653 | |a Zell- u. Molekularbiologie | ||
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| adam_text | Contents Preface to the Eighth Edition xv Part I The P,:-.ra65 Oonina arid PP P ••֊.•.і,֊ 1 1,3 1Д 1.5 1.6 The early development of genetics 4 The advent of gene cloning and the polymerase chain reaction 4 What is gene cloning? 5 What is PCR? 5 Why gene cloning and PCR are so important 8 1.5.1 Obtaining a pure sample of a gene by cloning 8 1.5.2 PCR can also be used to purify a gene 10 How to find your way through this book 11 Further reading ¡3 Vectors for Gene Cloning: Plasmids and Bacteriophages 15 2.1 2.2 3 j Why Gene Cloning and DIMA Analysis are Important 3 1.1 1 ֊2 2 vr- Pvsis Plasmids 15 2.1.1 Size and copy number 17 2.1.2 Conjugation and compatibility 18 2.1.3 Plasmid classification 19 2.1.4 Plasmids in organisms other than bacteria 19 Bacteriophages 19 2.2.1 The phage infection cycle 20 2.2.2 Lysogenic phages 20 2.2.3 Viruses as cloning vectors for other organisms 26 Further reading 27 Purification of DNA from Living Cells 29 3.1 Preparation of total cell DNA 30 3.1.1 Growing and harvesting a bacterial culture 30 3.1.2 Preparation of a cell extract 31
Contents 3.1.3 3.1.4 3.1.5 3.1.6 Purification of DNA from a cell extract ՝ ì Concentration of DNA samples г Measurement of DNA concentration ín Other methods for the preparation of total cell DNA 39 Preparation of plasmid DNA ֊to 3.2.1 Separation on the basis of size 4 ! 3.2.2 Separation on the basis of conformation 44 3.2.3 Plasmid amplification 44 Preparation of bacteriophage DNA 4 3.3.1 Growth of cultures to obtain a high / titre 4՜ 3.3.2 Preparation of non-lysogenic / pliages 4՜ 3.3.3 Collection of phages from an infected culture 49 3.3.4 Purification of DNA from λ phage particles 4լ 3.3.5 Purification of M13 DNA causes few problems 4և) Further reading 51 Manipulation of Purified DNA 51 4.1 The range of DNA manipulative enzymes 4.1.1 4.1.2 4.1.3 4.1.4 Nucleases 55 Ligases 5՜ Polymerases 5՜ DNA modifying enzymes 5S 4.2 Enzymes for cutting DNA - restriction endonucleases 59 4.2.1 The discovery and function of restriction endonucleases 60 4.2.2 Type II restriction endonucleases cut DNA at specific nucleotide sequences a I 4.2.3 Blunt ends and sticky ends 64 4.2.4 The frequency of recognition sequences in a DNA molecule 64 4.2.5 Performing a restriction digest in the laboratory 64 4.2.6 Analysing the result of restriction endonuclease cleavage 66 4.2.7 Estimation of the sizes of DNA molecules 68 4.2.8 Mapping the positions of different restriction sites in a DNA molecule 69 4.2.9 Special gel electrophoresis methods for separating larger molecules 70 4,3 Ligation - joining DNA molecules together 72 4.3.1 The mode of action of DNA ligase 72 4.3.2 Sticky ends increase the
efficiency of ligation 74 4.3.3 Putting sticky ends onto a blunt-ended molecule 74 4.3.4 Blunt-end ligation with a DNA topoisomerase 79 Further reading 81
ix Contents Introduction of DNA into Living Cells 85 Transformation - the uptake of DNA by bacterial cells 85 5.1.1 5.2 :5-3 Not all species of bacteria are equally efficient at DNA uptake 85 5.1.2 Preparation of competent E. coli cells 86 5.1.3 Selection for transformed cells 86 Identification of recombinants 88 5.2.1 Recombinant selection with pBR322 - insertional inactivation of an antibiotic resistance gene 89 5.2.2 Insertional inactivation does not always involve antibiotic resistance 90 Introduction of phage DNA into bacterial cells 9շ 5.3.1 5.3.2 5.3.3 Transfection 93 In vitro packaging of λ cloning vectors 95 Phage infection is visualized as plaques on an agar medium 92 5.3.4 Identification of recombinant phages 95 5,4 Introduction of DNA into non-bacterial cells 97 5.4.1 Transformation of individual cells 97 5.4.2 Transformation of whole organisms 99 Further reading 99 Cloning Vectors for E. coli 101 á. Cloning vectors based on E. coli plasmids 102 6.1.1 6.1.2 6.1.3 6.1.4 6.2 6.3 The nomenclature of plasmid cloning vectors 102 The useful properties of pBR322 102 The pedigree of pBR322 103 More sophisticated E. coli plasmid cloning vectors 104 Cloning vectors based on λ bacteriophage 108 6.2.1 Natural selection was used to isolate modified A that lack certain restriction sites 108 6.2.2 Segments of the λ genome can be deleted without impairing viability 108 6.2.3 Insertion and replacement vectors 110 6.2.4 Cloning experiments with A insertion or replacement vectors ¡12 6.2.5 Long DNA fragments can be cloned using a cosmid 113 6.2.6 A and other high-capacity
vectors enable genomic libraries to be constructed 114 Cloninq vectors for synthesis of sinqle-stranded DNA 115 6.3.1 6.3.2 Vectors based on M13 bacteriophage 1 15 Hybrid plasmid-M13 vectors 117
Contents x Vectors for other bacteria l l s Further reading l Ի Cloning Vectors for Eukaryotes І I і Vectors for yeast and other fungi 7.1.1 Selectable markers for the 2 μηι pi..smid IC2 7.1.2 Vectors based on the 2 pm plusm:cl yeast episomal plasmids ltd 7.1.3 A YEp may insert into yeast chromosomal DNA LM 7.1.4 Other types of yeast cloning erto: 7.1.5 Artificial chromosomes can be- us՛-M ;o clone long pieces of DNA in yeast 7.1.6 Vectors for other yeasts and fungi ՛ Cloning vectors for higher plants Լ՝ 7.2.1 Agrobacterium tumefaciens ֊ nature s smallest genetic engineer I ՝՛ 7.2.2 Cloning genes in plants by direct gene transfer I ՝5 7.2.3 Attempts to use plant viruses as cloning vectors І м Cloning vectors for animals I ;s 7.3.1 Cloning vectors for insects l ՝° 7.3.2 Cloning in mammals 1 11 Further reading 14.1 How to Obtain a Clone of a Specific Gene і 4 ՝ The problem of selection I4e 8.1.1 There are two basic strategies for obtaining the clone you want 14a Si Direct selection 14՜ 8.2.1 Marker rescue extends the scope of direct selection 144 8.2.2 The scope and limitations of marker rescue 150 8,3 Identification of a clone from a gene library 150 8.3.1 Gene libraries 151 H i Methods for clone identification 153 8.4.1 Complementary nucleic acid strands hybridize to each other 154 8.4.2 Colony and plaque hybridization probing 154 8.4.3 Examples of the practical use of hybridization probing 157 8.4.4 Identification methods based on detection of the translation product of the cloned gene 164 Further reading 166
Contents XI The Polymerase Chain Reaction l n t 9.1 PCR in outline 170 792 PCR in more detail I՜շ 9.2.1 9.2.2 9.3 Designing the oligonucleotide primers for a PCR 172 Working out the correct temperatures to use P I After the PCR: studying PCR products I n 9.3.1 9.3.2 9,A Gel electrophoresis of PCR products I Cloning PCR products I7,s Real-time PCR 180 9.4.1 9.4.2 9.4.3 Carrying out a real-time PCR experiment iso Real-time PCR enables the amount of starting material to be quantified 182 Melting curve analysis enables point mutations to be identified 184 Further reading 185 rart 11 The Appi · eternino з ri c! Dría ln 187 Sequencing Genes and Genomes 189 Chain-termination DNA sequencing 190 10.1.1 10.1.2 10.1.3 10.1.4 Chain-termination sequencing in outline 190 Not all DNA polymerases can be used for sequencing 192 Chain-termination sequencing with Taq polymerase 193 Limitations of chain-termination sequencing 195 Next-generation sequencing 196 10.2.1 10.2.2 10.2.3 10.2.4 10.2.5 10.2.6 10,3 How to 10.3.1 10.3.2 Preparing a library for an Illumina sequencing experiment 197 The sequencing phase of an Illumina experiment 199 Ion semiconductor sequencing 201 Third-generation sequencing 201 Next-generation sequencing without a DNA polymerase 202 Directing next-generation sequencing at specific sets of genes 203 sequence a genome 205 Shotgun sequencing of prokaryotic genomes 20o Sequencing of eukaryotic genomes 209 Further reading 215
xii Contents Studying Gene Expression and Function 2 l Studying the RNA transcript of a gene 2 1 ՝ 11.1.1 Detecting the presence or a transcript in an RNA sample 2 I4 11.1.2 Transcript mapping by hybridization between gene and RNA 22՚ 11.1.3 Transcript analysis by primer extension 222 11.1.4 Transcript analysis by PCR 22 Studying the regulation of gene expression 2 24 11.2.1 Identifying protein binding sites on a DNA molecule 224 Identifying control sequences by deletion analysis 24n Identifying and studying the translation product of a cloned gene 2 ΐ2 11.3.1 HRT and HART can identify the translation product of a cloned gene 2 ՝ ՝ 11.3.2 Analysis of proteins by in vitro mutagenesis 2 4 Further reading 240 11.2.2 Studying Genomes 244 12.2 12.3 Locating the genes in a genome sequence 2 N 12.1.1 Locating protein-coding genes by scanning a genome sequence 244 12.1.2 Gene location is aided by homology searching 24՜ 12.1.3 Locating genes for noncoding RNA transcripts 244 12.1.4 Identifying the binding sites for regulatory proteins in a genome sequence 2 7) Determining the function of an unknown gene 2՝ I 12.2.1 Assigning gene functions by computer analysis 241 12.2.2 Assigning gene function by experimental analysis 252 Genome browsers 256 Further reading 25T Studying Transcriptomes and Proteomes 254 13,1 Studying transcriptomes 259 13.1.1 Studying transcriptomes by microarray or chip analysis 260 13.1.2 Studying transcriptomes by RNA sequencing 2a l 13 2 Studying proteomes 265 13.2.1 Protein profiling 266 13.2.2 Studying protein-protein interactions 2~o Further reading 274
xiii Contents о* The Applications of Gene and DNA Anály d ՛՞է ·՝·֊--dnoloyy շ֊տ 14 Production of Protein from Cloned Genes 2 14.1 Special vectors for expression of foreign genes in E. coli 280 14.1.1 14.1.2 14.2 General problems with the production of recombinant protein in E. coli 287 14.2.1 14.2.2 14.3 The promoter is the critical component of an expression vector 28 I Cassettes and gene fusions 285 Problems resulting from the sequence of the foreign gene 288 Problems caused by E. coli 289 Production of recombinant protein by eukaryotic cells 290 14.3.1 Recombinant protein from yeast and filamentous fungi 291 14.3.2 Using animal cells for recombinant protein production 293 14.3.3 Pharming ֊ recombinant protein from live animals and plants 295 Further reading 298 15 Gene Cloning and DNA Analysis in Medicine ЗО і 15.1 Production of recombinant pharmaceuticals ЗО I 15.1.1 15.1.2 15.1.3 15.1.4 15.1.5 15.2 Identification of genes responsible for human diseases 314 15.2.1 15.2.2 15.3 Recombinant insulin 302 Synthesis of human growth hormones in E. coli 304 Recombinant factor VIII 305 Synthesis of other recombinant human proteins 308 Recombinant vaccines 308 How to identify a gene for a genetic disease 3I5 Genetic typing of disease mutations 320 Gene therapy 321 15.3.1 Gene therapy for inherited diseases 321 15.3.2 Gene therapy and cancer 325 15.3.3 The ethical issues raised by gene therapy 324 Further reading 325
Contents xiv ձ Gene Cloning and DNA Analysis in Agriculture 16.1 The gene addition approach to plant genetic engineering 12 s 16.1.1 Plants that make their own insecticides 4s 16.1.2 16.1.3 16.1.4 16.2 Herbicide-resistant crops ;՛՛■՛· Improving the nutritional quality or plants by gene addition ՝ 4 Other gene addition projects ՛ ՝s Gene subtraction 4m 16.2.1 16.3 16.4 Antisense RNA and the engine-m Ing of fruit ripening in tomato tin 16.2.2 Other examples of the use of antisense RNA in plant genetic engineering ; Gene editing with a programmable nuclease ill 16.3.1 Gene editing of phytoene desaturase iti rice 44 16.3.2 Editing of multiple genes in a single plant 4 6 16.3.3 Future developments in gene editing of plants 4՜ Are GM plants harmful to human health and the environment? 14 о 16.4.1 Safety concerns with selectable markers 4 о 16.4.2 The possibility of harmful effects on the environment їм) Further reading 24 і 17 Gene Cloning and DNA Analysis in Forensic Science and Archaeology 155 17.1 DNA analysis in the identification of crime suspects 44 17.1.1 17.2 17.1.2 Studying kinship by DNA profiling 15о 17.2.1 17.3 17.4 Genetic fingerprinting by hybridization probing 156 DNA profiling by PCR of short tandem repeats 15՜ Related individuals have similar DNA profiles 4ч 17.2.2 DNA profiling and the remains of the Romanovs 40 Sex identification by DNA analysis leli 17.3.1 PCRs directed at Y chromosome-specific sequences 361 17.3.2 PCR of the amelogenin gene 164 Archaeogenetics - using DNA to study human prehistory 365 17.4.1 The origins of modern humans .45 17.4.2 DNA can
also be used to study prehistoric human migrations 370 Further reading 374 Glossary 377 Index 395
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| adam_txt |
Contents Preface to the Eighth Edition xv Part I The P,:-.ra65 Oonina arid PP P ••֊.•.і,֊ 1 1,3 1Д 1.5 1.6 The early development of genetics 4 The advent of gene cloning and the polymerase chain reaction 4 What is gene cloning? 5 What is PCR? 5 Why gene cloning and PCR are so important 8 1.5.1 Obtaining a pure sample of a gene by cloning 8 1.5.2 PCR can also be used to purify a gene 10 How to find your way through this book 11 Further reading ¡3 Vectors for Gene Cloning: Plasmids and Bacteriophages 15 2.1 2.2 3 j Why Gene Cloning and DIMA Analysis are Important 3 1.1 1 ֊2 2 vr-'Pvsis Plasmids 15 2.1.1 Size and copy number 17 2.1.2 Conjugation and compatibility 18 2.1.3 Plasmid classification 19 2.1.4 Plasmids in organisms other than bacteria 19 Bacteriophages 19 2.2.1 The phage infection cycle 20 2.2.2 Lysogenic phages 20 2.2.3 Viruses as cloning vectors for other organisms 26 Further reading 27 Purification of DNA from Living Cells 29 3.1 Preparation of total cell DNA 30 3.1.1 Growing and harvesting a bacterial culture 30 3.1.2 Preparation of a cell extract 31
Contents 3.1.3 3.1.4 3.1.5 3.1.6 Purification of DNA from a cell extract ՝ ì Concentration of DNA samples г Measurement of DNA concentration ín Other methods for the preparation of total cell DNA 39 Preparation of plasmid DNA ֊to 3.2.1 Separation on the basis of size 4 ! 3.2.2 Separation on the basis of conformation 44 3.2.3 Plasmid amplification 44 Preparation of bacteriophage DNA 4 3.3.1 Growth of cultures to obtain a high / titre 4՜ 3.3.2 Preparation of non-lysogenic / pliages 4՜ 3.3.3 Collection of phages from an infected culture 49 3.3.4 Purification of DNA from λ phage particles 4լ 3.3.5 Purification of M13 DNA causes few problems 4և) Further reading 51 Manipulation of Purified DNA 51 4.1 The range of DNA manipulative enzymes 4.1.1 4.1.2 4.1.3 4.1.4 Nucleases 55 Ligases 5՜ Polymerases 5՜ DNA modifying enzymes 5S 4.2 Enzymes for cutting DNA - restriction endonucleases 59 4.2.1 The discovery and function of restriction endonucleases 60 4.2.2 Type II restriction endonucleases cut DNA at specific nucleotide sequences a I 4.2.3 Blunt ends and sticky ends 64 4.2.4 The frequency of recognition sequences in a DNA molecule 64 4.2.5 Performing a restriction digest in the laboratory 64 4.2.6 Analysing the result of restriction endonuclease cleavage 66 4.2.7 Estimation of the sizes of DNA molecules 68 4.2.8 Mapping the positions of different restriction sites in a DNA molecule 69 4.2.9 Special gel electrophoresis methods for separating larger molecules 70 4,3 Ligation - joining DNA molecules together 72 4.3.1 The mode of action of DNA ligase 72 4.3.2 Sticky ends increase the
efficiency of ligation 74 4.3.3 Putting sticky ends onto a blunt-ended molecule 74 4.3.4 Blunt-end ligation with a DNA topoisomerase 79 Further reading 81
ix Contents Introduction of DNA into Living Cells 85 Transformation - the uptake of DNA by bacterial cells 85 5.1.1 5.2 :5-3 Not all species of bacteria are equally efficient at DNA uptake 85 5.1.2 Preparation of competent E. coli cells 86 5.1.3 Selection for transformed cells 86 Identification of recombinants 88 5.2.1 Recombinant selection with pBR322 - insertional inactivation of an antibiotic resistance gene 89 5.2.2 Insertional inactivation does not always involve antibiotic resistance 90 Introduction of phage DNA into bacterial cells 9շ 5.3.1 5.3.2 5.3.3 Transfection 93 In vitro packaging of λ cloning vectors 95 Phage infection is visualized as plaques on an agar medium 92 5.3.4 Identification of recombinant phages 95 5,4 Introduction of DNA into non-bacterial cells 97 5.4.1 Transformation of individual cells 97 5.4.2 Transformation of whole organisms 99 Further reading 99 Cloning Vectors for E. coli 101 á. ' Cloning vectors based on E. coli plasmids 102 6.1.1 6.1.2 6.1.3 6.1.4 6.2 6.3 The nomenclature of plasmid cloning vectors 102 The useful properties of pBR322 102 The pedigree of pBR322 103 More sophisticated E. coli plasmid cloning vectors 104 Cloning vectors based on λ bacteriophage 108 6.2.1 Natural selection was used to isolate modified A that lack certain restriction sites 108 6.2.2 Segments of the λ genome can be deleted without impairing viability 108 6.2.3 Insertion and replacement vectors 110 6.2.4 Cloning experiments with A insertion or replacement vectors ¡12 6.2.5 Long DNA fragments can be cloned using a cosmid 113 6.2.6 A and other high-capacity
vectors enable genomic libraries to be constructed 114 Cloninq vectors for synthesis of sinqle-stranded DNA 115 6.3.1 6.3.2 Vectors based on M13 bacteriophage 1 15 Hybrid plasmid-M13 vectors 117
Contents x Vectors for other bacteria l l s Further reading l Ի Cloning Vectors for Eukaryotes І I і Vectors for yeast and other fungi 7.1.1 Selectable markers for the 2 μηι pi.smid IC2 7.1.2 Vectors based on the 2 pm plusm:cl yeast episomal plasmids ltd 7.1.3 A YEp may insert into yeast chromosomal DNA LM 7.1.4 Other types of yeast cloning \ erto: 7.1.5 Artificial chromosomes can be- us՛-M ;o clone long pieces of DNA in yeast 7.1.6 Vectors for other yeasts and fungi '՛ Cloning vectors for higher plants Լ՝ 7.2.1 Agrobacterium tumefaciens ֊ nature s smallest genetic engineer I'՝՛' 7.2.2 Cloning genes in plants by direct gene transfer I '՝5 7.2.3 Attempts to use plant viruses as cloning vectors І м Cloning vectors for animals I ;s 7.3.1 Cloning vectors for insects l ՝° 7.3.2 Cloning in mammals 1 11 Further reading 14.1 How to Obtain a Clone of a Specific Gene і 4 ՝ The problem of selection I4e 8.1.1 There are two basic strategies for obtaining the clone you want 14a Si Direct selection 14՜ 8.2.1 Marker rescue extends the scope of direct selection 144 8.2.2 The scope and limitations of marker rescue 150 8,3 Identification of a clone from a gene library 150 8.3.1 Gene libraries 151 H i Methods for clone identification 153 8.4.1 Complementary nucleic acid strands hybridize to each other 154 8.4.2 Colony and plaque hybridization probing 154 8.4.3 Examples of the practical use of hybridization probing 157 8.4.4 Identification methods based on detection of the translation product of the cloned gene 164 Further reading 166
Contents XI The Polymerase Chain Reaction l n't 9.1 PCR in outline 170 792 PCR in more detail I՜շ 9.2.1 9.2.2 9.3 Designing the oligonucleotide primers for a PCR 172 Working out the correct temperatures to use P I After the PCR: studying PCR products I n 9.3.1 9.3.2 9,A Gel electrophoresis of PCR products I Cloning PCR products I7,s Real-time PCR 180 9.4.1 9.4.2 9.4.3 Carrying out a real-time PCR experiment iso Real-time PCR enables the amount of starting material to be quantified 182 Melting curve analysis enables point mutations to be identified 184 Further reading 185 rart 11 The Appi'· eternino з ri c! Dría ln 187 Sequencing Genes and Genomes 189 Chain-termination DNA sequencing 190 10.1.1 10.1.2 10.1.3 10.1.4 Chain-termination sequencing in outline 190 Not all DNA polymerases can be used for sequencing 192 Chain-termination sequencing with Taq polymerase 193 Limitations of chain-termination sequencing 195 Next-generation sequencing 196 10.2.1 10.2.2 10.2.3 10.2.4 10.2.5 10.2.6 10,3 How to 10.3.1 10.3.2 Preparing a library for an Illumina sequencing experiment 197 The sequencing phase of an Illumina experiment 199 Ion semiconductor sequencing 201 Third-generation sequencing 201 Next-generation sequencing without a DNA polymerase 202 Directing next-generation sequencing at specific sets of genes 203 sequence a genome 205 Shotgun sequencing of prokaryotic genomes 20o Sequencing of eukaryotic genomes 209 Further reading 215
xii Contents Studying Gene Expression and Function 2 l Studying the RNA transcript of a gene 2 1 ՝ 11.1.1 Detecting the presence or a transcript in an RNA sample 2 I4 11.1.2 Transcript mapping by hybridization between gene and RNA 22՚ 11.1.3 Transcript analysis by primer extension 222 11.1.4 Transcript analysis by PCR 22' Studying the regulation of gene expression 2 24 11.2.1 Identifying protein binding sites on a DNA molecule 224 Identifying control sequences by deletion analysis 24n Identifying and studying the translation product of a cloned gene 2 ΐ2 11.3.1 HRT and HART can identify the translation product of a cloned gene 2 '՝ ՝ 11.3.2 Analysis of proteins by in vitro mutagenesis 2 4 Further reading 240 11.2.2 Studying Genomes 244 12.2 12.3 Locating the genes in a genome sequence 2 N 12.1.1 Locating protein-coding genes by scanning a genome sequence 244 12.1.2 Gene location is aided by homology searching 24՜ 12.1.3 Locating genes for noncoding RNA transcripts 244 12.1.4 Identifying the binding sites for regulatory proteins in a genome sequence 2 7) Determining the function of an unknown gene 2՝ I 12.2.1 Assigning gene functions by computer analysis 241 12.2.2 Assigning gene function by experimental analysis 252 Genome browsers 256 Further reading 25T Studying Transcriptomes and Proteomes 254 13,1 Studying transcriptomes 259 13.1.1 Studying transcriptomes by microarray or chip analysis 260 13.1.2 Studying transcriptomes by RNA sequencing 2a l 13 2 Studying proteomes 265 13.2.1 Protein profiling 266 13.2.2 Studying protein-protein interactions 2~o Further reading 274
xiii Contents о* The Applications of Gene and DNA Anály d ՛՞է ·՝·֊--dnoloyy շ֊տ 14 Production of Protein from Cloned Genes 2 14.1 Special vectors for expression of foreign genes in E. coli 280 14.1.1 14.1.2 14.2 General problems with the production of recombinant protein in E. coli 287 14.2.1 14.2.2 14.3 The promoter is the critical component of an expression vector 28 I Cassettes and gene fusions 285 Problems resulting from the sequence of the foreign gene 288 Problems caused by E. coli 289 Production of recombinant protein by eukaryotic cells 290 14.3.1 Recombinant protein from yeast and filamentous fungi 291 14.3.2 Using animal cells for recombinant protein production 293 14.3.3 Pharming ֊ recombinant protein from live animals and plants 295 Further reading 298 15 Gene Cloning and DNA Analysis in Medicine ЗО і 15.1 Production of recombinant pharmaceuticals ЗО I 15.1.1 15.1.2 15.1.3 15.1.4 15.1.5 15.2 Identification of genes responsible for human diseases 314 15.2.1 15.2.2 15.3 Recombinant insulin 302 Synthesis of human growth hormones in E. coli 304 Recombinant factor VIII 305 Synthesis of other recombinant human proteins 308 Recombinant vaccines 308 How to identify a gene for a genetic disease 3I5 Genetic typing of disease mutations 320 Gene therapy 321 15.3.1 Gene therapy for inherited diseases 321 15.3.2 Gene therapy and cancer 325 15.3.3 The ethical issues raised by gene therapy 324 Further reading 325
Contents xiv ձ Gene Cloning and DNA Analysis in Agriculture 16.1 The gene addition approach to plant genetic engineering 12 s 16.1.1 Plants that make their own insecticides 4s 16.1.2 16.1.3 16.1.4 16.2 ' Herbicide-resistant crops ;՛՛■՛· Improving the nutritional quality or plants by gene addition ՝ 4 Other gene addition projects ՛ ՝s Gene subtraction 4m 16.2.1 16.3 16.4 Antisense RNA and the engine-m Ing of fruit ripening in tomato tin 16.2.2 Other examples of the use of antisense RNA in plant genetic engineering ; Gene editing with a programmable nuclease ill 16.3.1 Gene editing of phytoene desaturase iti rice 44 16.3.2 Editing of multiple genes in a single plant 4 6 16.3.3 Future developments in gene editing of plants 4՜ Are GM plants harmful to human health and the environment? 14 о 16.4.1 Safety concerns with selectable markers 4 о 16.4.2 The possibility of harmful effects on the environment їм) Further reading 24 і 17 Gene Cloning and DNA Analysis in Forensic Science and Archaeology 155 17.1 DNA analysis in the identification of crime suspects 44 17.1.1 17.2 17.1.2 Studying kinship by DNA profiling 15о 17.2.1 17.3 17.4 Genetic fingerprinting by hybridization probing 156 DNA profiling by PCR of short tandem repeats 15՜ Related individuals have similar DNA profiles 4ч 17.2.2 DNA profiling and the remains of the Romanovs 40 Sex identification by DNA analysis leli 17.3.1 PCRs directed at Y chromosome-specific sequences 361 17.3.2 PCR of the amelogenin gene 164 Archaeogenetics - using DNA to study human prehistory 365 17.4.1 The origins of modern humans .45 17.4.2 DNA can
also be used to study prehistoric human migrations 370 Further reading 374 Glossary 377 Index 395 |
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| author | Brown, Terence A. 1953- |
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| genre | (DE-588)4151278-9 Einführung gnd-content |
| genre_facet | Einführung |
| id | DE-604.BV047081404 |
| illustrated | Illustrated |
| index_date | 2024-07-03T16:16:34Z |
| indexdate | 2024-07-10T09:02:03Z |
| institution | BVB |
| isbn | 9781119640783 |
| language | English |
| oai_aleph_id | oai:aleph.bib-bvb.de:BVB01-032488199 |
| oclc_num | 1227223128 |
| open_access_boolean | |
| owner | DE-355 DE-BY-UBR DE-703 DE-29T DE-19 DE-BY-UBM DE-11 |
| owner_facet | DE-355 DE-BY-UBR DE-703 DE-29T DE-19 DE-BY-UBM DE-11 |
| physical | xv, 416 Seiten Illustrationen, Diagramme, Karten |
| publishDate | 2021 |
| publishDateSearch | 2021 |
| publishDateSort | 2021 |
| publisher | Wiley Blackwell |
| record_format | marc |
| spelling | Brown, Terence A. 1953- Verfasser (DE-588)113251661 aut Gene cloning and DNA analysis an introduction T. A. Brown (University of Manchester, Manchester, United Kingdom) Eighth edition Hoboken, NJ ; Chichester Wiley Blackwell 2021 xv, 416 Seiten Illustrationen, Diagramme, Karten txt rdacontent n rdamedia nc rdacarrier Cloning, Molecular DNA, Recombinant - analysis Sequence Analysis, DNA Genklonierung (DE-588)4123275-6 gnd rswk-swf Sequenzanalyse Chemie (DE-588)4132277-0 gnd rswk-swf DNS (DE-588)4070512-2 gnd rswk-swf Biowissenschaften Life Sciences Genomforschung u. Proteomik Gentechnologie Genomics & Proteomics Genetik Genetics Cell & Molecular Biology Zell- u. Molekularbiologie DNS (DE-588)4151278-9 Einführung gnd-content Genklonierung (DE-588)4123275-6 s DE-604 DNS (DE-588)4070512-2 s Sequenzanalyse Chemie (DE-588)4132277-0 s Erscheint auch als Online-Ausgabe, PDF 978-1-119-64075-2 Erscheint auch als Online-Ausgabe, EPUB 978-1-119-64067-7 Digitalisierung UB Regensburg - ADAM Catalogue Enrichment application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=032488199&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Brown, Terence A. 1953- Gene cloning and DNA analysis an introduction Cloning, Molecular DNA, Recombinant - analysis Sequence Analysis, DNA Genklonierung (DE-588)4123275-6 gnd Sequenzanalyse Chemie (DE-588)4132277-0 gnd DNS (DE-588)4070512-2 gnd |
| subject_GND | (DE-588)4123275-6 (DE-588)4132277-0 (DE-588)4070512-2 (DE-588)4151278-9 |
| title | Gene cloning and DNA analysis an introduction |
| title_auth | Gene cloning and DNA analysis an introduction |
| title_exact_search | Gene cloning and DNA analysis an introduction |
| title_exact_search_txtP | Gene cloning and DNA analysis an introduction |
| title_full | Gene cloning and DNA analysis an introduction T. A. Brown (University of Manchester, Manchester, United Kingdom) |
| title_fullStr | Gene cloning and DNA analysis an introduction T. A. Brown (University of Manchester, Manchester, United Kingdom) |
| title_full_unstemmed | Gene cloning and DNA analysis an introduction T. A. Brown (University of Manchester, Manchester, United Kingdom) |
| title_short | Gene cloning and DNA analysis |
| title_sort | gene cloning and dna analysis an introduction |
| title_sub | an introduction |
| topic | Cloning, Molecular DNA, Recombinant - analysis Sequence Analysis, DNA Genklonierung (DE-588)4123275-6 gnd Sequenzanalyse Chemie (DE-588)4132277-0 gnd DNS (DE-588)4070512-2 gnd |
| topic_facet | Cloning, Molecular DNA, Recombinant - analysis Sequence Analysis, DNA Genklonierung Sequenzanalyse Chemie DNS Einführung |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=032488199&sequence=000001&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
| work_keys_str_mv | AT brownterencea genecloninganddnaanalysisanintroduction |