{"product_id":"gene-cloning-and-dna-analysis-9781119640783","title":"Gene Cloning and DNA Analysis","description":"\u003cb\u003eBook Synopsis\u003c\/b\u003e\u003cbr\u003e\u003cbr\u003e\u003cbr\u003e\u003cb\u003eTable of Contents\u003c\/b\u003e\u003cbr\u003e\u003cp\u003ePreface to the Eighth Edition xv\u003c\/p\u003e \u003cp\u003e\u003cb\u003ePart I The Basic Principles of Gene Cloning and DNA Analysis 1\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e1 Why Gene Cloning and DNA Analysis are Important 3\u003c\/p\u003e \u003cp\u003e2 Vectors for Gene Cloning: Plasmids and Bacteriophages 15\u003c\/p\u003e \u003cp\u003e3 Purification of DNA from Living Cells 29\u003c\/p\u003e \u003cp\u003e4 Manipulation of Purified DNA 53\u003c\/p\u003e \u003cp\u003e5 Introduction of DNA into Living Cells 83\u003c\/p\u003e \u003cp\u003e6 Cloning Vectors for \u003ci\u003eE. coli \u003c\/i\u003e101\u003c\/p\u003e \u003cp\u003e7 Cloning Vectors for Eukaryotes 121\u003c\/p\u003e \u003cp\u003e8 How to Obtain a Clone of a Specific Gene 145\u003c\/p\u003e \u003cp\u003e9 The Polymerase Chain Reaction 169\u003c\/p\u003e \u003cp\u003e\u003cb\u003ePart II The Applications of Gene Cloning and DNA Analysis in Research 187\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e10 Sequencing Genes and Genomes 189\u003c\/p\u003e \u003cp\u003e11 Studying Gene Expression and Function 217\u003c\/p\u003e \u003cp\u003e12 Studying Genomes 243\u003c\/p\u003e \u003cp\u003e13 Studying Transcriptomes and Proteomes 259\u003c\/p\u003e \u003cp\u003e\u003cb\u003ePart III The Applications of Gene Cloning and DNA Analysis in Biotechnology 275\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e14 Production of Protein from Cloned Genes 277\u003c\/p\u003e \u003cp\u003e15 Gene Cloning and DNA Analysis in Medicine 301\u003c\/p\u003e \u003cp\u003e16 Gene Cloning and DNA Analysis in Agriculture 327\u003c\/p\u003e \u003cp\u003e17 Gene Cloning and DNA Analysis in Forensic Science and Archaeology 355\u003c\/p\u003e \u003cp\u003eGlossary 377\u003c\/p\u003e \u003cp\u003eIndex 395\u003c\/p\u003e \u003cp\u003e \u003c\/p\u003e \u003cp\u003e \u003c\/p\u003e \u003cp\u003e \u003c\/p\u003e \u003cp\u003e \u003c\/p\u003e \u003cp\u003e \u003c\/p\u003e \u003cp\u003e \u003c\/p\u003e \u003cp\u003ePreface to the Eighth Edition xv\u003c\/p\u003e \u003cp\u003e\u003cb\u003ePart I The Basic Principles of Gene Cloning and DNA Analysis 1\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003cb\u003e1 Why Gene Cloning and DNA Analysis are Important 3\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e1.1 The early development of genetics 4\u003c\/p\u003e \u003cp\u003e1.2 The advent of gene cloning and the polymerase chain reaction 4\u003c\/p\u003e \u003cp\u003e1.3 What is gene cloning? 5\u003c\/p\u003e \u003cp\u003e1.4 What is PCR? 5\u003c\/p\u003e \u003cp\u003e1.5 Why gene cloning and PCR are so important 8\u003c\/p\u003e \u003cp\u003e1.5.1 Obtaining a pure sample of a gene by cloning 8\u003c\/p\u003e \u003cp\u003e1.5.2 PCR can also be used to purify a gene 10\u003c\/p\u003e \u003cp\u003e1.6 How to find your way through this book 11\u003c\/p\u003e \u003cp\u003eFurther reading 13\u003c\/p\u003e \u003cp\u003e\u003cb\u003e2 Vectors for Gene Cloning: Plasmids and Bacteriophages 15\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e2.1 Plasmids 15\u003c\/p\u003e \u003cp\u003e2.1.1 Size and copy number 17\u003c\/p\u003e \u003cp\u003e2.1.2 Conjugation and compatibility 18\u003c\/p\u003e \u003cp\u003e2.1.3 Plasmid classification 19\u003c\/p\u003e \u003cp\u003e2.1.4 Plasmids in organisms other than bacteria 19\u003c\/p\u003e \u003cp\u003e2.2 Bacteriophages 19\u003c\/p\u003e \u003cp\u003e2.2.1 The phage infection cycle 20\u003c\/p\u003e \u003cp\u003e2.2.2 Lysogenic phages 20\u003c\/p\u003e \u003cp\u003e2.2.3 Viruses as cloning vectors for other organisms 26\u003c\/p\u003e \u003cp\u003eFurther reading 27\u003c\/p\u003e \u003cp\u003e\u003cb\u003e3 Purification of DNA from Living Cells 29\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e3.1 Preparation of total cell DNA 30\u003c\/p\u003e \u003cp\u003e3.1.1 Growing and harvesting a bacterial culture 30\u003c\/p\u003e \u003cp\u003e3.1.2 Preparation of a cell extract 31\u003c\/p\u003e \u003cp\u003e3.1.3 Purification of DNA from a cell extract 33\u003c\/p\u003e \u003cp\u003e3.1.4 Concentration of DNA samples 37\u003c\/p\u003e \u003cp\u003e3.1.5 Measurement of DNA concentration 38\u003c\/p\u003e \u003cp\u003e3.1.6 Other methods for the preparation of total cell DNA 39\u003c\/p\u003e \u003cp\u003e3.2 Preparation of plasmid DNA 40\u003c\/p\u003e \u003cp\u003e3.2.1 Separation on the basis of size 41\u003c\/p\u003e \u003cp\u003e3.2.2 Separation on the basis of conformation 42\u003c\/p\u003e \u003cp\u003e3.2.3 Plasmid amplification 44\u003c\/p\u003e \u003cp\u003e3.3 Preparation of bacteriophage DNA 46\u003c\/p\u003e \u003cp\u003e3.3.1 Growth of cultures to obtain a high λ titre 47\u003c\/p\u003e \u003cp\u003e3.3.2 Preparation of non‐lysogenic λ phages 47\u003c\/p\u003e \u003cp\u003e3.3.3 Collection of phages from an infected culture 49\u003c\/p\u003e \u003cp\u003e3.3.4 Purification of DNA from λ phage particles 49\u003c\/p\u003e \u003cp\u003e3.3.5 Purification of M13 DNA causes few problems 49\u003c\/p\u003e \u003cp\u003eFurther reading 51\u003c\/p\u003e \u003cp\u003e\u003cb\u003e4 Manipulation of Purified DNA 53\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e4.1 The range of DNA manipulative enzymes 55\u003c\/p\u003e \u003cp\u003e4.1.1 Nucleases 55\u003c\/p\u003e \u003cp\u003e4.1.2 Ligases 57\u003c\/p\u003e \u003cp\u003e4.1.3 Polymerases 57\u003c\/p\u003e \u003cp\u003e4.1.4 DNA modifying enzymes 58\u003c\/p\u003e \u003cp\u003e4.2 Enzymes for cutting DNA – restriction endonucleases 59\u003c\/p\u003e \u003cp\u003e4.2.1 The discovery and function of restriction endonucleases 60\u003c\/p\u003e \u003cp\u003e4.2.2 Type II restriction endonucleases cut DNA at specific nucleotide sequences 61\u003c\/p\u003e \u003cp\u003e4.2.3 Blunt ends and sticky ends 62\u003c\/p\u003e \u003cp\u003e4.2.4 The frequency of recognition sequences in a DNA molecule 63\u003c\/p\u003e \u003cp\u003e4.2.5 Performing a restriction digest in the laboratory 64\u003c\/p\u003e \u003cp\u003e4.2.6 Analysing the result of restriction endonuclease cleavage 66\u003c\/p\u003e \u003cp\u003e4.2.7 Estimation of the sizes of DNA molecules 68\u003c\/p\u003e \u003cp\u003e4.2.8 Mapping the positions of different restriction sites in a DNA molecule 69\u003c\/p\u003e \u003cp\u003e4.2.9 Special gel electrophoresis methods for separating larger molecules 70\u003c\/p\u003e \u003cp\u003e4.3 Ligation – joining DNA molecules together 72\u003c\/p\u003e \u003cp\u003e4.3.1 The mode of action of DNA ligase 72\u003c\/p\u003e \u003cp\u003e4.3.2 Sticky ends increase the efficiency of ligation 74\u003c\/p\u003e \u003cp\u003e4.3.3 Putting sticky ends onto a blunt‐ended molecule 74\u003c\/p\u003e \u003cp\u003e4.3.4 Blunt‐end ligation with a DNA topoisomerase 79\u003c\/p\u003e \u003cp\u003eFurther reading 81\u003c\/p\u003e \u003cp\u003e\u003cb\u003e5 Introduction of DNA into Living Cells 83\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e5.1 Transformation – the uptake of DNA by bacterial cells 85\u003c\/p\u003e \u003cp\u003e5.1.1 Not all species of bacteria are equally efficient at DNA uptake 85\u003c\/p\u003e \u003cp\u003e5.1.2 Preparation of competent \u003ci\u003eE. coli \u003c\/i\u003ecells 86\u003c\/p\u003e \u003cp\u003e5.1.3 Selection for transformed cells 86\u003c\/p\u003e \u003cp\u003e5.2 Identification of recombinants 88\u003c\/p\u003e \u003cp\u003e5.2.1 Recombinant selection with pBR322 – insertional inactivation of an antibiotic resistance gene 89\u003c\/p\u003e \u003cp\u003e5.2.2 Insertional inactivation does not always involve antibiotic resistance 90\u003c\/p\u003e \u003cp\u003e5.3 Introduction of phage DNA into bacterial cells 92\u003c\/p\u003e \u003cp\u003e5.3.1 Transfection 93\u003c\/p\u003e \u003cp\u003e5.3.2 \u003ci\u003eIn vitro \u003c\/i\u003epackaging of λ cloning vectors 93\u003c\/p\u003e \u003cp\u003e5.3.3 Phage infection is visualized as plaques on an agar medium 93\u003c\/p\u003e \u003cp\u003e5.3.4 Identification of recombinant phages 95\u003c\/p\u003e \u003cp\u003e5.4 Introduction of DNA into non‐bacterial cells 97\u003c\/p\u003e \u003cp\u003e5.4.1 Transformation of individual cells 97\u003c\/p\u003e \u003cp\u003e5.4.2 Transformation of whole organisms 99\u003c\/p\u003e \u003cp\u003eFurther reading 99\u003c\/p\u003e \u003cp\u003e\u003cb\u003e6 Cloning Vectors for \u003ci\u003eE. coli \u003c\/i\u003e101\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e6.1 Cloning vectors based on \u003ci\u003eE. coli \u003c\/i\u003eplasmids 102\u003c\/p\u003e \u003cp\u003e6.1.1 The nomenclature of plasmid cloning vectors 102\u003c\/p\u003e \u003cp\u003e6.1.2 The useful properties of pBR322 102\u003c\/p\u003e \u003cp\u003e6.1.3 The pedigree of pBR322 103\u003c\/p\u003e \u003cp\u003e6.1.4 More sophisticated \u003ci\u003eE. coli \u003c\/i\u003eplasmid cloning vectors 104\u003c\/p\u003e \u003cp\u003e6.2 Cloning vectors based on \u003cb\u003eλ \u003c\/b\u003ebacteriophage 108\u003c\/p\u003e \u003cp\u003e6.2.1 Natural selection was used to isolate modified \u003cb\u003e\u003ci\u003eλ \u003c\/i\u003e\u003c\/b\u003ethat lack certain restriction sites 108\u003c\/p\u003e \u003cp\u003e6.2.2 Segments of the \u003cb\u003e\u003ci\u003eλ \u003c\/i\u003e\u003c\/b\u003egenome can be deleted without impairing viability 108\u003c\/p\u003e \u003cp\u003e6.2.3 Insertion and replacement vectors 110\u003c\/p\u003e \u003cp\u003e6.2.4 Cloning experiments with \u003cb\u003e\u003ci\u003eλ \u003c\/i\u003e\u003c\/b\u003einsertion or replacement vectors 112\u003c\/p\u003e \u003cp\u003e6.2.5 Long DNA fragments can be cloned using a cosmid 113\u003c\/p\u003e \u003cp\u003e6.2.6 \u003cb\u003e\u003ci\u003eλ \u003c\/i\u003e\u003c\/b\u003eand other high‐capacity vectors enable genomic libraries to be constructed 114\u003c\/p\u003e \u003cp\u003e6.3 Cloning vectors for synthesis of single‐stranded DNA 115\u003c\/p\u003e \u003cp\u003e6.3.1 Vectors based on M13 bacteriophage 115\u003c\/p\u003e \u003cp\u003e6.3.2 Hybrid plasmid–M13 vectors 117\u003c\/p\u003e \u003cp\u003e6.4 Vectors for other bacteria 118\u003c\/p\u003e \u003cp\u003eFurther reading 119\u003c\/p\u003e \u003cp\u003e\u003cb\u003e7 Cloning Vectors for Eukaryotes 121\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e7.1 Vectors for yeast and other fungi 121\u003c\/p\u003e \u003cp\u003e7.1.1 Selectable markers for the 2 μm plasmid 122\u003c\/p\u003e \u003cp\u003e7.1.2 Vectors based on the 2 μm plasmid – yeast episomal plasmids 122\u003c\/p\u003e \u003cp\u003e7.1.3 A YEp may insert into yeast chromosomal DNA 124\u003c\/p\u003e \u003cp\u003e7.1.4 Other types of yeast cloning vector 124\u003c\/p\u003e \u003cp\u003e7.1.5 Artificial chromosomes can be used to clone long pieces of DNA in yeast 126\u003c\/p\u003e \u003cp\u003e7.1.6 Vectors for other yeasts and fungi 129\u003c\/p\u003e \u003cp\u003e7.2 Cloning vectors for higher plants 129\u003c\/p\u003e \u003cp\u003e7.2.1 \u003ci\u003eAgrobacterium tumefaciens \u003c\/i\u003e– nature’s smallest genetic engineer 130\u003c\/p\u003e \u003cp\u003e7.2.2 Cloning genes in plants by direct gene transfer 135\u003c\/p\u003e \u003cp\u003e7.2.3 Attempts to use plant viruses as cloning vectors 137\u003c\/p\u003e \u003cp\u003e7.3 Cloning vectors for animals 138\u003c\/p\u003e \u003cp\u003e7.3.1 Cloning vectors for insects 139\u003c\/p\u003e \u003cp\u003e7.3.2 Cloning in mammals 141\u003c\/p\u003e \u003cp\u003eFurther reading 143\u003c\/p\u003e \u003cp\u003e\u003cb\u003e8 How to Obtain a Clone of a Specific Gene 145\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e8.1 The problem of selection 146\u003c\/p\u003e \u003cp\u003e8.1.1 There are two basic strategies for obtaining the clone you want 146\u003c\/p\u003e \u003cp\u003e8.2 Direct selection 147\u003c\/p\u003e \u003cp\u003e8.2.1 Marker rescue extends the scope of direct selection 149\u003c\/p\u003e \u003cp\u003e8.2.2 The scope and limitations of marker rescue 150\u003c\/p\u003e \u003cp\u003e8.3 Identification of a clone from a gene library 150\u003c\/p\u003e \u003cp\u003e8.3.1 Gene libraries 151\u003c\/p\u003e \u003cp\u003e8.4 Methods for clone identification 153\u003c\/p\u003e \u003cp\u003e8.4.1 Complementary nucleic acid strands hybridize to each other 154\u003c\/p\u003e \u003cp\u003e8.4.2 Colony and plaque hybridization probing 154\u003c\/p\u003e \u003cp\u003e8.4.3 Examples of the practical use of hybridization probing 157\u003c\/p\u003e \u003cp\u003e8.4.4 Identification methods based on detection of the translation product of the cloned gene 164\u003c\/p\u003e \u003cp\u003eFurther reading 166\u003c\/p\u003e \u003cp\u003e\u003cb\u003e9 The Polymerase Chain Reaction 169\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e9.1 PCR in outline 170\u003c\/p\u003e \u003cp\u003e9.2 PCR in more detail 172\u003c\/p\u003e \u003cp\u003e9.2.1 Designing the oligonucleotide primers for a PCR 172\u003c\/p\u003e \u003cp\u003e9.2.2 Working out the correct temperatures to use 174\u003c\/p\u003e \u003cp\u003e9.3 After the PCR: studying PCR products 176\u003c\/p\u003e \u003cp\u003e9.3.1 Gel electrophoresis of PCR products 177\u003c\/p\u003e \u003cp\u003e9.3.2 Cloning PCR products 178\u003c\/p\u003e \u003cp\u003e9.4 Real‐time PCR 180\u003c\/p\u003e \u003cp\u003e9.4.1 Carrying out a real‐time PCR experiment 180\u003c\/p\u003e \u003cp\u003e9.4.2 Real‐time PCR enables the amount of starting material to be quantified 182\u003c\/p\u003e \u003cp\u003e9.4.3 Melting curve analysis enables point mutations to be identified 184\u003c\/p\u003e \u003cp\u003eFurther reading 185\u003c\/p\u003e \u003cp\u003e\u003cb\u003ePart II The Applications of Gene Cloning and DNA Analysis in Research 187\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003cb\u003e10 Sequencing Genes and Genomes 189\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e10.1 Chain‐termination DNA sequencing 190\u003c\/p\u003e \u003cp\u003e10.1.1 Chain‐termination sequencing in outline 190\u003c\/p\u003e \u003cp\u003e10.1.2 Not all DNA polymerases can be used for sequencing 192\u003c\/p\u003e \u003cp\u003e10.1.3 Chain‐termination sequencing with \u003ci\u003eTaq \u003c\/i\u003epolymerase 193\u003c\/p\u003e \u003cp\u003e10.1.4 Limitations of chain‐termination sequencing 195\u003c\/p\u003e \u003cp\u003e10.2 Next‐generation sequencing 196\u003c\/p\u003e \u003cp\u003e10.2.1 Preparing a library for an Illumina sequencing experiment 197\u003c\/p\u003e \u003cp\u003e10.2.2 The sequencing phase of an Illumina experiment 199\u003c\/p\u003e \u003cp\u003e10.2.3 Ion semiconductor sequencing 201\u003c\/p\u003e \u003cp\u003e10.2.4 Third‐generation sequencing 201\u003c\/p\u003e \u003cp\u003e10.2.5 Next‐generation sequencing without a DNA polymerase 202\u003c\/p\u003e \u003cp\u003e10.2.6 Directing next‐generation sequencing at specific sets of genes 203\u003c\/p\u003e \u003cp\u003e10.3 How to sequence a genome 205\u003c\/p\u003e \u003cp\u003e10.3.1 Shotgun sequencing of prokaryotic genomes 206\u003c\/p\u003e \u003cp\u003e10.3.2 Sequencing of eukaryotic genomes 209\u003c\/p\u003e \u003cp\u003eFurther reading 215\u003c\/p\u003e \u003cp\u003e\u003cb\u003e11 Studying Gene Expression and Function 217\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e11.1 Studying the RNA transcript of a gene 218\u003c\/p\u003e \u003cp\u003e11.1.1 Detecting the presence of a transcript in an RNA sample 219\u003c\/p\u003e \u003cp\u003e11.1.2 Transcript mapping by hybridization between gene and RNA 220\u003c\/p\u003e \u003cp\u003e11.1.3 Transcript analysis by primer extension 222\u003c\/p\u003e \u003cp\u003e11.1.4 Transcript analysis by PCR 223\u003c\/p\u003e \u003cp\u003e11.2 Studying the regulation of gene expression 224\u003c\/p\u003e \u003cp\u003e11.2.1 Identifying protein binding sites on a DNA molecule 225\u003c\/p\u003e \u003cp\u003e11.2.2 Identifying control sequences by deletion analysis 230\u003c\/p\u003e \u003cp\u003e11.3 Identifying and studying the translation product of a cloned gene 232\u003c\/p\u003e \u003cp\u003e11.3.1 HRT and HART can identify the translation product of a cloned gene 233\u003c\/p\u003e \u003cp\u003e11.3.2 Analysis of proteins by \u003ci\u003ein vitro \u003c\/i\u003emutagenesis 234\u003c\/p\u003e \u003cp\u003eFurther reading 240\u003c\/p\u003e \u003cp\u003e\u003cb\u003e12 Studying Genomes 243\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e12.1 Locating the genes in a genome sequence 244\u003c\/p\u003e \u003cp\u003e12.1.1 Locating protein‐coding genes by scanning a genome sequence 244\u003c\/p\u003e \u003cp\u003e12.1.2 Gene location is aided by homology searching 247\u003c\/p\u003e \u003cp\u003e12.1.3 Locating genes for noncoding RNA transcripts 249\u003c\/p\u003e \u003cp\u003e12.1.4 Identifying the binding sites for regulatory proteins in a genome sequence 250\u003c\/p\u003e \u003cp\u003e12.2 Determining the function of an unknown gene 251\u003c\/p\u003e \u003cp\u003e12.2.1 Assigning gene functions by computer analysis 251\u003c\/p\u003e \u003cp\u003e12.2.2 Assigning gene function by experimental analysis 252\u003c\/p\u003e \u003cp\u003e12.3 Genome browsers 256\u003c\/p\u003e \u003cp\u003eFurther reading 257\u003c\/p\u003e \u003cp\u003e\u003cb\u003e13 Studying Transcriptomes and Proteomes 259\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e13.1 Studying transcriptomes 259\u003c\/p\u003e \u003cp\u003e13.1.1 Studying transcriptomes by microarray or chip analysis 260\u003c\/p\u003e \u003cp\u003e13.1.2 Studying transcriptomes by RNA sequencing 261\u003c\/p\u003e \u003cp\u003e13.2 Studying proteomes 265\u003c\/p\u003e \u003cp\u003e13.2.1 Protein profiling 266\u003c\/p\u003e \u003cp\u003e13.2.2 Studying protein–protein interactions 270\u003c\/p\u003e \u003cp\u003eFurther reading 274\u003c\/p\u003e \u003cp\u003e\u003cb\u003ePart III The Applications of Gene Cloning and DNA Analysis in Biotechnology 275\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e\u003cb\u003e14 Production of Protein from Cloned Genes 277\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e14.1 Special vectors for expression of foreign genes in \u003ci\u003eE. coli \u003c\/i\u003e280\u003c\/p\u003e \u003cp\u003e14.1.1 The promoter is the critical component of an expression vector 281\u003c\/p\u003e \u003cp\u003e14.1.2 Cassettes and gene fusions 285\u003c\/p\u003e \u003cp\u003e14.2 General problems with the production of recombinant protein in \u003ci\u003eE. coli \u003c\/i\u003e287\u003c\/p\u003e \u003cp\u003e14.2.1 Problems resulting from the sequence of the foreign gene 288\u003c\/p\u003e \u003cp\u003e14.2.2 Problems caused by \u003ci\u003eE. coli \u003c\/i\u003e289\u003c\/p\u003e \u003cp\u003e14.3 Production of recombinant protein by eukaryotic cells 290\u003c\/p\u003e \u003cp\u003e14.3.1 Recombinant protein from yeast and filamentous fungi 291\u003c\/p\u003e \u003cp\u003e14.3.2 Using animal cells for recombinant protein production 293\u003c\/p\u003e \u003cp\u003e14.3.3 Pharming – recombinant protein from live animals and plants 295\u003c\/p\u003e \u003cp\u003eFurther reading 298\u003c\/p\u003e \u003cp\u003e\u003cb\u003e15 Gene Cloning and DNA Analysis in Medicine 301\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e15.1 Production of recombinant pharmaceuticals 301\u003c\/p\u003e \u003cp\u003e15.1.1 Recombinant insulin 302\u003c\/p\u003e \u003cp\u003e15.1.2 Synthesis of human growth hormones in \u003ci\u003eE. coli \u003c\/i\u003e304\u003c\/p\u003e \u003cp\u003e15.1.3 Recombinant factor VIII 305\u003c\/p\u003e \u003cp\u003e15.1.4 Synthesis of other recombinant human proteins 308\u003c\/p\u003e \u003cp\u003e15.1.5 Recombinant vaccines 308\u003c\/p\u003e \u003cp\u003e15.2 Identification of genes responsible for human diseases 314\u003c\/p\u003e \u003cp\u003e15.2.1 How to identify a gene for a genetic disease 315\u003c\/p\u003e \u003cp\u003e15.2.2 Genetic typing of disease mutations 320\u003c\/p\u003e \u003cp\u003e15.3 Gene therapy 321\u003c\/p\u003e \u003cp\u003e15.3.1 Gene therapy for inherited diseases 321\u003c\/p\u003e \u003cp\u003e15.3.2 Gene therapy and cancer 323\u003c\/p\u003e \u003cp\u003e15.3.3 The ethical issues raised by gene therapy 324\u003c\/p\u003e \u003cp\u003eFurther reading 325\u003c\/p\u003e \u003cp\u003e\u003cb\u003e16 Gene Cloning and DNA Analysis in Agriculture 327\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e16.1 The gene addition approach to plant genetic engineering 328\u003c\/p\u003e \u003cp\u003e16.1.1 Plants that make their own insecticides 328\u003c\/p\u003e \u003cp\u003e16.1.2 Herbicide‐resistant crops 334\u003c\/p\u003e \u003cp\u003e16.1.3 Improving the nutritional quality of plants by gene addition 337\u003c\/p\u003e \u003cp\u003e16.1.4 Other gene addition projects 338\u003c\/p\u003e \u003cp\u003e16.2 Gene subtraction 339\u003c\/p\u003e \u003cp\u003e16.2.1 Antisense RNA and the engineering of fruit ripening in tomato 340\u003c\/p\u003e \u003cp\u003e16.2.2 Other examples of the use of antisense RNA in plant genetic engineering 342\u003c\/p\u003e \u003cp\u003e16.3 Gene editing with a programmable nuclease 344\u003c\/p\u003e \u003cp\u003e16.3.1 Gene editing of phytoene desaturase in rice 344\u003c\/p\u003e \u003cp\u003e16.3.2 Editing of multiple genes in a single plant 346\u003c\/p\u003e \u003cp\u003e16.3.3 Future developments in gene editing of plants 347\u003c\/p\u003e \u003cp\u003e16.4 Are GM plants harmful to human health and the environment? 349\u003c\/p\u003e \u003cp\u003e16.4.1 Safety concerns with selectable markers 349\u003c\/p\u003e \u003cp\u003e16.4.2 The possibility of harmful effects on the environment 350\u003c\/p\u003e \u003cp\u003eFurther reading 351\u003c\/p\u003e \u003cp\u003e\u003cb\u003e17 Gene Cloning and DNA Analysis in Forensic Science and Archaeology 355\u003c\/b\u003e\u003c\/p\u003e \u003cp\u003e17.1 DNA analysis in the identification of crime suspects 356\u003c\/p\u003e \u003cp\u003e17.1.1 Genetic fingerprinting by hybridization probing 356\u003c\/p\u003e \u003cp\u003e17.1.2 DNA profiling by PCR of short tandem repeats 357\u003c\/p\u003e \u003cp\u003e17.2 Studying kinship by DNA profiling 359\u003c\/p\u003e \u003cp\u003e17.2.1 Related individuals have similar DNA profiles 359\u003c\/p\u003e \u003cp\u003e17.2.2 DNA profiling and the remains of the Romanovs 360\u003c\/p\u003e \u003cp\u003e17.3 Sex identification by DNA analysis 363\u003c\/p\u003e \u003cp\u003e17.3.1 PCRs directed at Y chromosome‐specific sequences 363\u003c\/p\u003e \u003cp\u003e17.3.2 PCR of the amelogenin gene 364\u003c\/p\u003e \u003cp\u003e17.4 Archaeogenetics – using DNA to study human prehistory 365\u003c\/p\u003e \u003cp\u003e17.4.1 The origins of modern humans 365\u003c\/p\u003e \u003cp\u003e17.4.2 DNA can also be used to study prehistoric human migrations 370\u003c\/p\u003e \u003cp\u003eFurther reading 374\u003c\/p\u003e \u003cp\u003eGlossary 377\u003c\/p\u003e \u003cp\u003eIndex 395\u003c\/p\u003e","brand":"John Wiley and Sons 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