Note: Intro -- Contents -- Introduction -- 1 Actinomycetes and Antibiotics -- 2 Antibiotic Discovery and Resistance -- 3 Microbial Sex -- 4 Toward Gene Cloning -- 5 From Chromosome Map to DNA Sequence -- 6 Bacteria That Develop -- 7 The Switch to Antibiotic Production -- 8 Unnatural Natural Products -- 9 Functional Genomics -- 10 Genomics Against Tuberculosis and Leprosy -- Conclusion -- Notes and References -- Glossary -- A -- B -- C -- D -- E -- F -- G -- H -- I -- K -- L -- M -- N -- O -- P -- R -- S -- T -- U -- V -- Index -- A -- B -- C -- D -- E -- F -- G -- H -- I -- J -- K -- L -- M -- N -- O -- P -- Q -- R -- S -- T -- U -- V -- W -- X -- Z.

Note: Front Cover -- Natural Product Biosynthesis by Microorganisms and Plants, Part C -- Copyright -- Contents -- Contributors -- Preface -- Methods in Enzymology -- Section 1: Metabolites from Novel Sources -- Chapter 1: Methods for the Study of Endophytic Microorganisms from Traditional Chinese Medicine Plants -- 1. Introduction -- 2. Experimental Methods -- 2.1. Medicinal plant selection and sampling -- 2.2. Pretreatment and surface-sterilization -- 2.2.1. Pretreatment -- 2.2.2. Surface sterilization -- 2.3. Isolation of endophytes -- Media for isolation of endophytic bacteria and actinobacteria -- Media for isolation of endophytic fungi -- 2.4. Purification and maintenance -- 2.5. Taxonomic analysis -- 3. Novel Bioactive Compounds from Endophytic Actinobacteria -- 4. Conclusion -- Acknowledgment -- References -- Chapter 2: Cyanobacteria as a Source of Natural Products -- 1. Introduction -- 2. Cyanobacteria as Starting Material for Natural Products -- 2.1. Isolation of cyanobacterial strains -- 2.2. Mass cultivation of cyanobacterial strains -- 2.2.1. Protocol for mass cultivation of cyanobacteria in a laboratory scale -- 2.3. Harvesting cyanobacteria from natural habitats -- 2.3.1. Protocol: Harvesting cyanobacterial cells from field habitats -- 3. Screening for New Compounds and Producers -- 3.1. Screening for new compounds -- 3.1.1. Protocol: Screening by whole-cell mass spectrometry of freeze-dried cyanobacterial biomass for the presence of mic ... -- 3.2. Characterization of (new?) natural products -- 3.3. Finding new sources of known compounds -- 4. Genomic Mining of Cyanobacterial Natural Products -- 4.1. Mutagenesis of biosynthetic genes in cyanobacteria -- 4.1.1. Genetic manipulation of cyanobacteria -- 4.2. Transcriptional activation of biosynthetic genes in cyanobacteria. , 4.3. Heterologous production of natural products from cyanobacteria -- 4.4. In vitro reconstitution of cyanobacterial biosynthetic pathways -- 5. Summary -- References -- Chapter 3: Isolating Antifungals from Fungus-Growing Ant Symbionts Using a Genome-Guided Chemistry Approach -- 1. Introduction -- 2. Strain Isolation and Genome Sequencing -- 2.1. Selectively isolating actinomycetes -- 2.2. Bioassaying for antifungal activity -- 2.3. Genome sequencing and analysis of Pseudonocardia P1 and S. albus S4 -- 3. Identifying Antifungals made by Pseudonocardia P1 -- 3.1. Searching for antifungal pathways in the P1 genome -- 3.2. Chemical identity of nystatin P1 -- 3.3. Attempts at heterologous expression -- 4. Identifying Antifungals made by S. albus S4 -- 4.1. Searching for antifungal pathways in the S4 genome -- 4.2. Disrupting candicidin biosynthesis -- 4.3. Identification of the antimycin gene cluster -- 4.4. Cloning the antimycin gene cluster -- 4.5. Other gene clusters in the S4 genome -- 5. Summary and Perspectives -- Acknowledgments -- References -- Chapter 4: Gamma-Butyrolactone and Furan Signaling Systems in Streptomyces -- 1. Introduction -- 2. AfsA-Like Enzymes and the Biosynthesis of Signaling Molecules -- 2.1. A-factor biosynthesis by AfsA -- 2.2. The role of MmfL in MMF biosynthesis -- 3. Heterologous Expression of an AfsA-Like-Containing Mini-Cluster Combined with Comparative Metabolic Profiling -- 4. Genetic Disruption of mmfLHP and Chemical Complementation Experiments -- 5. Conclusion -- Acknowledgment -- References -- Chapter 5: Old Meets New: Using Interspecies Interactions to Detect Secondary Metabolite Production in Actinomycetes -- 1. Introduction -- 2. Setting Up Binary Actinomycete Interactions -- 2.1. Actinomycete isolation medium -- 2.2. Desiccation of soil -- 2.3. Generation of spore stocks for long-term storage of actinomycetes. , 2.4. Calculation and setup of interaction arrays -- 3. Activity-Guided Fractionation for Amychelin Isolation -- 3.1. Biological assays to guide purification of small molecules -- 3.2. Large-scale fermentation of AA4 and activity-guided fractionation -- 3.3. Generation and purification of Ga- or Fe-amychelin complexes -- 4. Determining the Affinity of Amychelin for Fe -- 5. Identifying the Amychelin Biosynthetic Gene Cluster -- 6. Transcriptomic Analysis using Nanostring Technology -- 7. Summary -- Acknowledgments -- References -- Section 2: Analyzing Gene Clusters -- Chapter 6: Finding and Analyzing Plant Metabolic Gene Clusters -- 1. Introduction -- 2. Discovery of Plant Metabolic Gene Clusters Through Genetic Analysis -- 2.1. Discovery of secondary metabolic clusters for the synthesis of defense compounds in maize and oat -- 2.2. Example: identification of the avenacin cluster in oat -- 3. Mining Genomes for Secondary Metabolic Gene Clusters -- 3.1. Predicting candidate metabolic gene clusters -- 3.2. Defining the scaffold -- 3.3. Example: analysis of an oxidosqualene cyclase by expression in yeast -- 4. Validating Candidate Metabolic Gene Clusters -- 4.1. Heterologous expression approaches -- 4.2. Reverse genetic approaches -- 4.3. Metabolite identification -- 4.4. Example: analysis of knockouts and knockdowns for the thalianol gene cluster -- 5. Imaging Gene Clusters -- 5.1. RNA probe preparation -- 5.2. DNA probe preparation -- 5.3. Preparation of sections for RNA and DNA fluorescence in situ hybridization (FISH) -- 5.4. RNA in situ hybridization and posthybridization washes -- 5.4.1. Solutions -- 5.5. DNA in situ hybridization and posthybridization washes -- 5.6. Immunodetection -- 5.7. Imaging -- 6. Future Prospects -- Acknowledgments -- References -- Chapter 7: Genomic Approaches for Interrogating the Biochemistry of Medicinal Plant Species. , 1. Introduction -- 2. Plant Material Selection and Quality -- 2.1. Germplasm selection -- 2.2. Plant health -- 2.3. Tissue selection -- 3. Isolation of RNA -- 4. Generation of Next-Generation whole Transcriptome Sequences -- 4.1. Removal of contaminating DNA in the RNA samples -- 4.2. Construction of cDNA libraries for next-generation sequencing -- 4.3. Sequencing of cDNA libraries -- 5. Assembly of a Reference Transcriptome -- 5.1. Quality assessment of sequences and cleaning of reads -- 5.2. De novo assembly of reads -- 5.3. Quality of the de novo assembly -- 5.4. Mapping of reads from single-tissue libraries -- 5.5. Improved de novo assembly -- 5.6. Quality assessment and functional annotation of the final assembly -- 6. Assessment of Expression Abundances -- 6.1. Mapping of reads to final de novo assembly -- 6.2. Quantification of the transcript abundances -- 6.3. Quality assessment of the expression matrix -- 7. Summary -- 8. Useful Links -- Acknowledgments -- References -- Chapter 8: Phylogenetic Approaches to Natural Product Structure Prediction -- 1. Introduction -- 1.1. A short introduction to phylogeny -- 1.2. The biosynthetic logic of secondary metabolism -- 1.2.1. Polyketide synthases -- 1.2.2. Nonribosomal peptide synthetases -- 1.2.3. More examples -- 2. Working with Sequence Data -- 2.1. Assembling the dataset -- 2.2. Creating alignments -- 2.3. Editing the alignment -- 2.4. Model tests -- 2.5. Generating phylogenetic trees -- 2.6. Bioinformatic programs -- 3. NaPDoS -- 3.1. Scope of NaDoS -- 3.2. How NaPDoS works -- 4. Conclusions and Future Directions -- Acknowledgments -- References -- Section 3: Heterologous Expression of Pathways -- Chapter 9: Using a Virus-Derived System to Manipulate Plant Natural Product Biosynthetic Pathways -- 1. Introduction -- 2. Deleted Vectors Based on Cowpea Mosaic Virus. , 2.1. Replication-competent deleted vectors -- 2.2. The CPMV-HT expression system -- 2.3. The pEAQ series of vectors -- 3. Expression of Enzymes in Plants Using pEAQ Vectors -- 3.1. Materials -- 3.1.1. Media, buffers, and solutions -- 3.1.2. Bacterial strains -- 3.1.3. Plants -- 3.2. Creating expression plasmids -- 3.2.1. Choice of expression vector -- 3.2.2. Restriction enzyme-based cloning -- 3.2.3. Cloning by GATEWAY recombination -- 3.2.4. Transformation of Agrobacterium -- 3.3. Agroinfiltration of N. benthamiana -- 3.3.1. Preparation of Agrobacterium suspensions -- 3.3.2. Infiltration of leaves with Agrobacterium suspensions -- 3.4. Monitoring expression levels in N. benthamiana leaves -- 4. Conclusions and Perspectives -- Acknowledgments -- References -- Chapter 10: DNA Assembler: A Synthetic Biology Tool for Characterizing and Engineering Natural Product Gene Clusters -- 1. Introduction -- 2. General Guidelines for Construct Design -- 3. Heterologous Expression and Fine Modification of Natural Product Gene Clusters -- 3.1. Heterologous expression of the aureothin biosynthetic gene cluster -- 3.1.1. DNA preparation -- 3.1.2. Transformation -- 3.1.3. Verification of the correctly assembled pathways -- 3.1.4. Conjugation and heterologous expression of the aureothin pathway variants in S. lividans -- 3.1.5. Detection of aureothin (and its derivatives) -- 3.2. Site-directed mutagenesis for generating new derivatives -- 3.2.1. DNA preparation -- 3.2.2. Detection and analysis of the aureothin derivatives -- 4. Characterization of Natural Product Gene Clusters -- 4.1. Heterologous expression of the spectinabilin biosynthetic gene cluster -- 4.1.1. Construction of the spectinabilin biosynthetic gene cluster -- 4.1.2. Detection of spectinabilin -- 4.2. Gene inactivation and scar-less gene deletion -- 5. Construction of Artificial Gene Clusters. , 5.1. Creation of a hybrid spectinabilin-aureothin gene cluster.

Note: Front Cover -- Methods in Enzymology Complex Enzymes in Microbial Natural Product Biosynthesis, Part B: Polyketides, Aminocoumarins and Carbohydrates -- Copyright Page -- Contents -- Contributors -- Preface -- Methods in Enzymology -- Chapter 1: Introduction to Polyketide Biosynthesis -- 1. Introduction -- 1.1. Types of PKS -- 1.2. Type II PKS -- 1.3. Type III PKS -- 1.4. Type I PKS -- 1.5. Combinatorial biosynthesis: Prospects and progress -- Acknowledgments -- References -- Chapter 2: Structural Enzymology of polyketide Synthases -- Chapter 3: Chapter Three: Fungal Type I Polyketide Synthases -- 1. Introduction -- 2. Partially Reducing PKSs: 6-Methylsalicylate Synthase -- 3. Nonreducing PKSs -- 3.1. Norsolorinic acid synthase -- 3.2. Tetrahydroxynaphthalene synthase -- 3.3. Bikaverin nonaketide synthase -- 4. Highly Reducing PKSs -- 4.1. Lovastatin (LNKS and LDKS) -- 4.2. HR PKS-NRPS: Fusarin and tenellin synthetases -- 5. NR/HR PKS Hybrid Systems: Zearalenone (ZAE1 and ZAE2) -- 6. Conclusions -- References -- Chapter 4: Tandem Acyl Carrier Protein Domains in Polyunsaturated Fatty Acid Synthases -- 1. Introduction -- 2. Methods -- 2.1. Production of PUFAs in E. coli by expressing the PUFAS genes -- 2.2. Mapping the active sites of PfaA-ACPs by site-directed mutagenesis -- 2.3. Overproduction of each of the PfaA-ACPs -- 2.4. Overproduction of PfaE and Svp PPTases -- 2.5. In vivo and in vitro preparation of the holo-form of PfaA-ACPs -- 2.6. Elucidation of the relationship between PUFA production and the number of active ACPs -- 3. Conclusion -- Acknowledgments -- References -- Chapter 5: Iterative Type I Polyketide Synthases for Enediyne Core Biosynthesis -- 1. Introduction -- 2. Methods -- 2.1. PCR amplification of PKSE cassettes for predictive classification of new enediynes -- 2.2. Heterologous expression and overproduction of PKSE proteins. , 2.3. Production and isolation of the polyene intermediate from 9-membered PKSEs -- 2.4. Production of apo-ACPs from PKSE for in vitro functional analyses -- 2.5. In vitro preparation of holo-ACPs -- 3. Conclusion -- Acknowledgments -- References -- Chapter 6: Chapter Six The DEBS Paradigm for Type I Modular Polyketide Synthases and Beyond -- 1. Introduction -- 2. DEBS and the Concept of a Module -- 2.1. Generalizability of the DEBS paradigm -- 3. Beyond the DEBS Paradigm -- 3.1. Specificity of the AT domains -- 3.2. Novel loading modules -- 3.3. Methylation domains -- 3.4. Trans PKS activities -- 3.5. Unusual modular organization -- 3.6. Unusual module functions -- 3.7. Intermodular interactions -- 4. Conclusion -- References -- Chapter 7: Formation and Characterization of Acyl Carrier Protein-Linked Polyketide Synthase Extender Units -- 1. Introduction -- 2. Overproduction and Purification of Recombinant Proteins -- 2.1. Principle -- 2.2. Materials -- 2.3. Heterologous overproduction of proteins -- 2.4. Purification of enzymes using batch-binding method with Nickel-NTA resin -- 3. Formation and Characterization of Hydroxymalonyl-ACP and Aminomalonyl-ACP -- 3.1. In vitro phosphopantetheinylation of the ACPs ZmaD and ZmaH -- 3.2. HPLC-based characterization of modified ACPs -- 3.3. Formation of (2R)-hydroxymalonyl-ACP -- 3.4. Formation of (2S)-aminomalonyl-ACP -- 3.5. MALDI-TOF MS analysis of ACPs -- 3.6. Other methods for characterizing enzymes involved in (2S)-aminomalonyl-ACP formation -- 3.7. Other methods for characterizing enzymes involved in (2R)-hydroxymalonyl-ACP formation -- References -- Chapter 8: Type I Polyketide Synthases That Require Discrete Acyltransferases -- 1. Introduction -- 2. Methods -- 2.1. Heterologous expression and overproduction of apo-ACPs from AT-less PKS modules -- 2.2. In vitro preparation of holo-ACPs. , 2.3. Heterologous expression and overproduction of discrete ATs -- 2.4. In vitro assay for AT substrate specificity -- 2.5. In vitro assay of AT-catalyzed loading of acyl CoA extender substrate to holo-ACPs -- 3. Conclusion -- Acknowledgments -- References -- Chapter 9: The Enzymology of Polyether Biosynthesis -- 1. Introduction -- 2. Genetic Mining and Functional Analysis of Genes Specific to Polyether Biosynthetic Pathways -- 3. Premonensin, the Parent Unsaturated Monensin Polyketide -- 4. Epoxidases MonCI, NigCI, and NanO -- 5. Epoxide Hydrolases MonBI/BII, NigBI/BII, NanI, and Lsd19 -- 6. NanE, a Polyether-Specific Thioesterase -- 6.1. In vitro studies of NanE thioesterase -- 7. Assay of Polyether Thioesterase Activity -- 7.1. Site-directed mutagenesis of NanE -- 8. Transcriptional Analysis of the Nanchangmycin Biosynthetic Pathway Genes -- 9. Conclusions -- Acknowledgments -- References -- Chapter 10: Enzymology of the Polyenes Pimaricin and Candicidin Biosynthesis -- 1. Introduction -- 2. Pimaricin as a Prototype of Small Polyenes: Discovery and Properties -- 3. Pimaricin Biosynthesis in S. natalensis -- 3.1. The pimaricin gene cluster -- 3.2. Formation of pimaricinolide: The pimaricinolide synthase complex -- 3.3. Pimaricinolide tailoring and export -- 4. Regulation of Pimaricin Biosynthesis -- 4.1. Transcriptional regulators -- 4.2. Regulation by cholesterol oxidase -- 4.3. Inducers of pimaricin biosynthesis -- 4.4. Global regulatory mechanisms -- 5. Candicidin: A Prototype of ``Aromatic´´ Polyenes -- 6. The Candicidin/FR-008 Gene Cluster -- 7. Biosynthesis of PABA: The pabAB and pabC Genes -- 8. The Polyketide Synthases -- 9. Monooxygenase Genes: Modifications of the Polyketide Chain -- 10. Transporter Genes -- 11. Genes Related to Mycosamine Biosynthesis -- 12. Regulatory Genes -- 13. Phosphate Represses Expression of the pabAB Gene. , 14. Future Perspectives -- Acknowledgments -- References -- Chapter 11: Genetic Analysis of Nystatin and Amphotericin Biosynthesis -- 1. Introduction -- 2. Gene Inactivation and Replacement in the Nystatin Producer Streptomyces noursei -- 2.1. Conjugative transfer of a recombinant plasmid from E. coli ET12567 (pUZ8002) into S. noursei ATCC 11455 -- 2.2. Gene inactivation in S. noursei -- 2.3. Gene replacement in S. noursei -- 3. Gene Inactivation and Replacement in the Amphotericin Producer Streptomyces nodosus -- 3.1. Phage-mediated gene replacement in S. nodosus -- 4. Production, Purification, and Characterization of Novel Amphotericin- and Nystatin-Related Polyenes -- 4.1. Production and identification of nystatin-related polyenes -- 4.2. Scaled-up production of nystatin analogues -- 4.3. Preparative LC-MS purification of nystatin analogues -- 5. Conclusion -- Acknowledgments -- References -- Chapter 12: Polyketide Versatility in the Biosynthesis of Complex Mycobacterial Cell Wall Lipids -- 1. Introduction -- 2. Acetate and Propionate Feeding Studies -- 3. Genome Sequencing and Identification of Polyketide Synthases -- 4. Mycobacterial Polyketide Synthases -- 4.1. Biosynthesis of dimycocerosate esters (DIMs) by PKS15/1, PpsABCDE, and MAS -- 4.2. PKS2 is involved in biosynthesis of sulfolipids -- 4.3. PKS12 uses a novel ``modularly iterative´´ mechanism for biosynthesis of mannosyl-beta-1-phosphomycoketides -- 4.4. PKS13 catalyzes condensation of fatty-acyl chains during biosynthesis of mycolic acids -- 4.5. PKS3/4 is involved in the biosynthesis of phthenoic acids -- 4.6. PKS10, PKS7, PKS8, PKS17, PKS9, and PKS11 constitute an unusual PKS cluster -- 4.7. PKS18 is involved in biosynthesis of long-chain pyrones -- 4.8. MbtC and MbtD are involved in biosynthesis of iron-chelating siderophores from Mtb -- 4.9. PKS5 and PKS6. , 5.1. Product formation assay to study activity of PKS enzymes -- 5.2. Characterization of PKS derived saturated fatty acids -- References -- Chapter 13: Genetic Engineering to Produce Polyketide Analogues -- 1. Introduction -- 2. AT Domain Replacement to Alter α-Carbon Substitution -- 3. Procedure for Engineering AT Replacements in the Chromosome -- 4. Engineering beta-Carbon Processing -- 5. Engineering when only a Single Crossover Event is Possible -- 6. Heterologous Expression of Engineered PKS Genes -- 7. Chemobiosynthesis -- 8. Mutasynthesis -- 9. Gene Knockouts to Obtain Analogues -- References -- Chapter 14: Design and Synthesis of Pathway Genes for Polyketide Biosynthesis -- 1. Introduction -- 2. Redesign of PKS Genes to Accommodate Unique Restriction Sites Flanking Individual Components and for Efficient Expression in E -- 3. Validation of Synthetic PKS Gene Design -- 4. A Rapid Assay to Identify Productive Combinations of PKS Modules -- 5. Assembly of Larger Polyketide Synthases Using Information Gained with the Bimodular Assay -- 6. Design and Construction of Synthetic Operons for the Expression of Sugar Pathway Genes -- References -- Chapter 15: Heterologous Production of Polyketides in Bacteria -- 1. Introduction -- 2. General Considerations for the Heterologous Expression of Polyketide Pathways -- 3. S. coelicolor as a Model System for Heterologous Expression of Polyketides -- 4. Procedure for the Heterologous Production of Polyketides in S. coelicolor -- 5. System Improvements for the Heterologous Production of Polyketides in Streptomyces spp. -- 5.1. Handling large PKS genes -- 5.2. Improving polyketide titers -- 5.3. Optimization of conjugation protocols for industrial strains -- 5.4. Optimizing polyketide precursors supply -- 6. Recent Developments for the Production of Polyketides in Nonactinomycete Bacteria. , 6.1. E. coli as heterologous host.

Note: Front Cover -- Natural Product Biosynthesis by Microorganisms and Plants, Part A -- Copyright -- Contents -- Contributors -- Preface -- Methods in Enzymology -- Section1:Terpenoids -- Chapter 1:Steady-State Kinetic Characterization of Sesquiterpene Synthases by Gas Chromatography-Mass Spectroscopy -- 1. Introduction -- 1.1. The vial assay for enzyme kinetics -- 1.2. Steady-state kinetics and the Michaelis-Menten model -- 2. Experimental Components and Considerations -- 2.1. Enzyme purification and quantification -- 2.2. Substrates -- 2.3. Buffer and pH -- 2.4. Metal ions -- 2.5. Internal standard -- 2.6. Vial assay method -- 2.7. GC-MS instrument and run parameters -- 2.8. Additional points -- 3. Pilot Experiments -- 3.1. Analyte detection and quantification -- 3.2. Instrument calibration -- 3.3. Linear range of protein concentration: Measuring kcat apparent -- 4. Steady-State Kinetic Experiments -- 4.1. Reaction velocity versus substrate concentration -- 4.2. Steady-state kinetics experiment -- 5. Data Handling/Processing -- 6. Summary -- References -- Chapter 2: Automating Gene Library Synthesis by Structure-Based Combinatorial Protein Engineering: Examples from Plant Sesquiterpene Synthses -- 1. Overview -- 2. Plasmid Library Synthesis -- 2.1. Deconstructing the gene -- 2.2. Cloning of the three-plasmid system -- 3. Optimization of SCOPE PCR Conditions -- 4. Generic SCOPE Method -- 4.1. Primer design -- 4.2. Fragment amplification -- 4.3. SCOPE synthesis: Recombination -- 4.4. SCOPE synthesis: Amplification -- 5. Applications of SCOPE Synthesis -- 5.1. Synthesis of complex mixtures of diverse mutants -- 5.1.1. Deconstructing the gene -- 5.1.2. Plasmid library synthesis -- 5.1.3. SCOPE synthesis -- 5.2. Synthesis of arrays of individual mutants -- 5.2.1. Deconstructing the gene -- 5.2.2. Plasmid library synthesis and fragment amplification. , 5.2.3. SCOPE synthesis -- 6. Troubleshooting -- 6.1. Presence of alternative amplification products -- 6.2. Low levels of full-length gene product -- 7. Conclusions -- References -- Chapter 3:In Planta Transient Expression Analysis of Monoterpene Synthases -- 1. Introduction -- 2. Plant Transformation Vector Construction -- 2.1. Materials -- 2.2. Preparing attB-PCR products using attB adaptor PCR -- 2.3. Protocol for destination vector cloning -- 3. Agrobacterium-Mediated Transient Expression -- 3.1. Materials -- 3.2. Preparation of competent A. tumefaciens -- 3.3. Transformation of A. tumefaciens GV3101 -- 3.4. Preparation of transformed Agrobacterium for transient expression -- 3.5. Transient transformation of N. benthamiana leaves -- 4. Agrobacterium Preparation for High-Throughput Analysis -- 5. Product Identification -- 5.1. Headspace volatile analysis -- 6. Transient Expression of a Kiwifruit Linalool Synthase -- 6.1. Vectors and cloning -- 6.2. Infiltration setup -- 6.3. Solvent extraction and analysis -- 6.4. Headspace trapping and analysis -- 6.5. Results and discussion -- 7. Conclusions -- Acknowledgments -- References -- Chapter 4:Natural Rubber Biosynthesis in Plants: Rubber Transferase -- 1. Introduction -- 2. Preparing Samples for Assaying Rubber Transferase Activity -- 2.1. Rubber particle-bound activity -- 2.1.1. Purification of enzymatically active rubber particles -- 2.1.2. Enzyme stability -- 2.1.3. Assay system purity -- 2.1.4. Sources of competing enzymes and the separate roles of cis and trans prenyl transferases in rubber biosynthesis -- 3. The Rubber Transferase Assay -- 3.1. Assay method -- 3.2. Kinetic analyses -- 3.3. Regulation of molecular weight -- 4. Identification and Purification of Rubber Transferase -- 4.1. Rubber transferase is a distinct cis prenyl transferase -- 4.2. Photoaffinity labeling. , 4.2.1. A photoaffinity labeling protocol -- 4.2.2. Purification of labeled proteins -- 4.3. Antibodies -- 4.3.1. Immunoinhibition and immunoprecipitation -- 4.3.2. Latex-free animals -- 5. Qualitative Protein Analysis -- 6. Protein Quantification -- 7. Summary -- Acknowledgments -- References -- Chapter 5:Discovery and Characterization of Terpenoid Biosynthetic Pathways of Fungi -- 1. Introduction -- 2. Identification, Cloning, and Characterization of Terpene Synthases -- 2.1. Identification and cloning of terpene synthase genes without sequence information -- 2.2. Identification and cloning of terpene synthase genes using sequence information -- 2.3. Characterization of terpene synthase gene function -- 3. Structural Characterization and Engineering of Fungal Terpene Synthase Activities -- 3.1. Site-directed mutagenesis guided by crystal structures -- 3.2. Exploring terpene synthase structure through molecular models -- 4. Identification and Characterization of Biosynthetic Gene Clusters -- 4.1. Characteristics of fungal biosynthetic gene clusters -- 4.2. Obtaining a biosynthetic cluster sequence -- 4.2.1. Identifying an anchor gene sequence -- 4.2.2. Obtaining a sequencing dataset -- 4.2.2.1. Small-scale sequencing approaches -- 4.2.2.2. Whole-genome sequencing -- 4.3. Cluster annotation -- 4.4. Characterization of biosynthetic clusters -- 4.4.1. Gene knockout and complementation studies -- 4.4.2. Pathway construction in a heterologous host -- 5. Genome Mining for Sesquiterpene Synthases -- 5.1. Phylogeny-guided analysis of terpene synthase gene families -- 6. Conclusions -- Acknowledgments -- References -- Chapter 6:Menaquinone Biosyntheses in Microorganisms -- 1. Introduction -- 2. Futalosine Pathway -- 2.1. Tracer experiments clearly showed the presence of an alternative pathway -- 2.2. Genes participating in the alternative pathway. , 2.3. Intermediates in the alternative pathway -- 2.3.1. Isolation of an intermediate accumulated by the SCO4327-disruptant -- 2.3.2. Identification of the intermediate next to FL -- 2.3.3. Identification of the intermediate following DHFL -- 2.3.4. Identification of the intermediate following cyclic DHFL -- 2.4. The early biosynthetic steps -- Acknowledgments -- References -- Chapter 7:Diversity and Analysis of Bacterial Terpene Synthases -- 1. Terpenoid Metabolites from Bacterial Cultures -- 2. Bacterial Terpene Synthases -- 2.1. Monoterpene synthases -- 2.2. Sesquiterpene synthases -- 2.3. Diterpene synthases -- 3. Methods for the Study of Bacterial Terpene Synthases -- 3.1. Bioinformatic analysis of bacterial terpene synthases -- 3.1.1. Properties of terpene synthases and database search procedures -- 3.1.2. Genome mining of bacterial terpene synthases -- 3.2. Expression of genes encoding bacterial terpene synthases in heterologous hosts -- Protocol 1. Construction of expression cassette on integrating vector for terpene synthase gene of interest -- Protocol 2. Introduction of recombinant integrating plasmid into S. avermitilis SUKA17 -- Protocol 3. Cultivation for the production of terpene compounds and analysis of products -- References -- Chapter 8: Platensimycin and Platencin Biosynthesis in Streptomyces platensis, Showcasing Discovery and Characterization of Novel Bacteria Diterpene Synthases -- 1. Introduction -- 2. Methods -- 2.1. In vivo confirmation of PtmT1 and PtmT3 as DTSs in PTM and PTN biosynthesis -- 2.1.1. Bioinformatic analyses to identify candidate DTSs -- 2.1.2. Construction of a gene replacement or deletion on an isolated cosmid -- 2.1.3. Replacement of the DTS gene in vivo with the antibiotic resistance cassette -- 2.1.4. Southern analysis to verify the mutant strain genotype. , 2.1.5. HPLC analysis to determine the mutant strain chemotype -- 2.2. In vivo confirmation of ptmT3 encoding a DTS by heterologous expression -- 2.2.1. Construction of heterologous expression strain -- 2.2.2. Structural validation of the diterpenoids produced in heterologous hosts -- 2.3. In vitro characterization of PtmT2 and PtmT3 as DTSs in PTM biosynthesis -- 2.3.1. Expression, overproduction, and purification of PtmT2 and PtmT3 from E. coli -- 2.3.2. Synthesis of GGDP from geranylgeraniol -- 2.3.3. Functional characterization of PtmT2 as an ent-CDP synthase -- 2.3.4. Functional characterization of PtmT3 as an ent-kauran-16-ol synthase -- 3. Conclusions -- Acknowledgment -- References -- Section 2:Alkaloids andGlucosinolates -- Section 2: Alkaloids and Glucosinolates -- Chapter 9:Strategies for Engineering Plant Natural Products: The Iridoid-Derived Monoterpene Indole Alkaloids of Catharanthus roseus -- 1. Introduction -- 2. Metabolic Engineering Strategies -- 2.1. Precursor-directed biosynthesis -- 2.1.1. Example of directed biosynthesis of monoterpene indole alkaloids -- 2.1.2. Methods for precursor-directed biosynthesis in C. roseus seedlings and hairy root cultures -- 2.2. Mutasynthesis -- 2.2.1. Example of mutasynthesis for monoterpene indole alkaloids -- 2.2.2. Methods for mutasynthesis in C. roseus -- 2.3. Incorporation of engineered pathway enzymes in biosynthesis -- 2.3.1. Example of incorporation of an engineered pathway enzyme in C. roseus -- 2.3.2. Methods for incorporation of an engineered pathway enzyme in C. roseus -- 3. Conclusions and Future Directions -- References -- Chapter 10:Discovery and Functional Analysis of Monoterpenoid Indole Alkaloid Pathways in Plants -- 1. Introduction -- 2. Enzyme Assays for MIA Pathway Steps -- 2.1. Tabersonine-16-hydroxylase assay coupled to 16-OMT -- 2.2. Tabersonine 6,7-epoxidase assay. , 2.3. Tabersonine 19 hydroxylase assay.

Note: Front Cover -- Genetics of Bacterial Diversity -- Copyright Page -- Table of Contents -- Contributors -- Preface -- Section I: Introductory Chapters: The Diversity of Bacteria and of Bacterial Genetics -- Chapter 1. Bacterial Diversity: The Range of Interesting Things that Bacteria Do -- I. Introduction -- II. Ecological Niches -- III. Towards a Phylogeny of Bacteria -- IV. Reinvention Throughout the Phylogenetic Tree -- V. Some Unexpected Attributes of Bacteria -- VI. Conclusion -- Acknowledgements -- References -- Chapter 2. Diversity of Bacterial Genetics -- I. Introduction -- II. The Prokaryotic Genome -- III. Transfer of Chromosomal DNA between Bacteria -- IV. Gene Expression -- V. Gene-, Pathway- and Regulon-specific Regulatory Mechanisms -- VI. Differences between Prokaryote and Eukaryote Genetics -- VII. Closing Remarks -- Acknowledgements -- References -- Chapter 3. Cloning and Molecular Analysis of Bacterial Genes -- I. Introduction -- II. Cloning Bacterial DNA -- III. Mutagenesis with Cloned DNA -- IV. Biochemical Procedures that Exploit Cloned DNA -- V. Current Limitations and Possibilities -- References -- Section II: Specialized Metabolic Capabilities of Bacteria -- Chapter 4. Regulation of Luminescence in Marine Bacteria -- I. Introduction -- II. Organization and Function of lux Genes -- III. Regulation of lux Expression -- IV. Luminescence Variation -- V. Conclusions -- Acknowledgements -- References -- Chapter 5. Photosynthesis in Rhodospirillaceae -- I. Introduction -- II. Structure-Function of the Photosynthetic Apparatus -- III. Protein Components of the Photosynthetic Apparatus -- IV. Photosynthetic Apparatus Genes -- V. In Vitro Mutagenesis Studies -- VI. Genetic Engineering in Reaction Centres -- References -- Chapter 6. The Genetics of Nitrogen Fixation -- I. The Diversity of Nitrogen-Fixing Bacteria. , II. The nif Genes of Klebsiellapneumoniae -- III. The Assembly of Active Nitrogenase -- IV. The Biochemistry and Physiology of Nitrogenase -- V. The Three Nitrogenases of Azotobacter -- VI. nif Genes in Other Organisms -- VII. Rearrangement of nif Genes in Anabaena -- VIII. Regulation of Expression oï nif Genes -- IX. Concluding Remarks -- Acknowledgements -- References -- Chapter 7. Antibiotic Biosynthesis in Streptomyces -- I. Introduction to Streptomyces Biology -- II. Antibiotic Production -- III. Molecular Genetics of Antibiotic Production -- IV. Overview, Implications and Prospects -- References -- Chapter 8. Catabolism of Aromatic Hydrocarbons by Pseudomonas -- I. Introduction -- II. Biochemical Strategies for Oxidative Catabolism of Aromatics -- III. Organization and Regulation of Genes for Catabolism of Aromatic Hydrocarbons -- IV. Utility of Determinants of Catabolic Pathways -- V. Laboratory Evolution of Aromatic Catabolic Pathways -- VI. Concluding remarks -- Acknowledgements -- References -- Chapter 9. Mercury Resistance in Bacteria -- I. Introduction -- II. Bacterial Transformations of Mercury -- III. Mercury Resistance Genes -- IV. The Gram-Negative Structural Genes and their Products -- V. A Model for Mercury Resistance in Bacteria -- VI. Regulation of Expression of the Mercury Resistance Genes -- VII. Overview and Prospects -- Acknowledgements -- References -- Section III: Morphological Differentiation-Flagella Spores and Multicellular Development -- Chapter 10. Differentiation in Caulobacter: Flagellum Development, Mobility and Chemotaxis -- I. Introduction -- II. Developmental Programmes and Cell Differentiation -- III. Regulation of Flagellum Biosynthesis -- IV. Control of Chemotaxis and Positioning of Differentiated Structures -- V. Prospects-The Cell Cycle as a Regulator of Temporal and Spatial Patterning -- Acknowledgements. , References -- Chapter 11. Pathways of Developmentally Regulated Gene Expression in Bacillus subtilis -- I. Introduction -- II. Sporulation and Germination -- III. Genes Involved in Sporulation and Germination -- IV. Developmental Genes are Switched on in an Ordered Temporal Sequence -- V. Compartmentalization of Gene Expression -- VI. Dependence Patterns of Developmental Gene Expression: Four Examples -- VII. Pathways of Developmentally Regulated Gene Expression -- VIII. Overview, Implications and Prospects -- Acknowledgements -- References -- Chapter 12. Multicellular Development in Myxobacteria -- I. Introduction -- II. Fruiting Body Development Follows a Programme -- III. Operon Fusions Expose a Programme of Differential Gene Expression . -- IV. Cell Interactions Coordinate the Programme of Fruiting Body Development -- V. Mutants of Groups A, B, C and D Differ Genetically -- VI. Expression of β Galactosidase from lac Fusion Strains Depends on the Products of the asg, bsg, csg and dsg Genes -- VII. A-factor and C-factor Activities can be Found in Cell Extracts -- VIII. The asg, bsg, csg and dsg Loci can be Isolated -- IX. Overview and Prospects -- References -- Section IV: Bacterial Adaptations to Animal Pathogenicity -- Chapter 13. The Molecular Basis ofAntigenic Variation in Pathogenic Neisseria -- I. Introduction -- II. Diversity and Virulence -- III. Genetic Mechanisms for Pilus Variation -- IV. Genetic Mechanisms for P.II Variation -- V. Conclusions -- Acknowledgements -- References -- Chapter 14. Adhesins of Pathogenic Escherichia coli -- I. Introduction -- II. Bacterial Adherence to Animal Tissues -- III. Adhesin Genetics -- IV. Evolutionary Perspectives -- References -- Chapter 15. Genetic Studies of Enterotoxin and Other Potential Virulence Factors q/'Vibrio cholerae -- I. Introduction -- II. Cholera Toxin Genes. , III. Adherence and Colonization -- IV. Other Potential Virulence Factors -- V. Regulation of Virulence Gene Expression -- VI. Perspectives -- References -- Chapter 16. Iron Scavenging in the Pathogenesis o/Escherichia coli -- I. Introduction -- II. Enterobacterial Iron Uptake Systems -- III. Molecular Genetics -- IV. Biochemical Genetics -- V. Regulation -- VI. Epilogue -- References -- Section V: Bacteria that Interact with Plants as Parasites or Symbionts -- Chapter 17. Pathogenicity of Xanthomonas and Related Bacteria Towards Plants -- I. Introduction -- II. Strategies and Techniques for Studying the Genetics of Pathogenicity -- III. Function of Some Pathogenicity Genes -- IV. Concluding Remarks -- References -- Chapter 18. Tumorigenicity of Agrobacterium on Plants -- I. Introduction -- II. Ti and Ri Plasmids -- III. T-DNA -- IV. Genes and Sequences Necessary for T-DNA Transfer -- V. Different Steps in the Process of Tumour Induction -- VI. Prospects for Application -- References -- Chapter 19. The Symbiosis Between Rhizobium and Legumes -- I. Introduction -- II. Methods for Identifying Bacterial Genes Involved in Nodulation -- III. Polysaccharide Synthesis is Important for Nodulation -- IV. Analysis oinod Gene Function -- V. Regulation of nod Gene Transcription -- VI. Conclusions -- References -- Section VI: Bacterial Population Genetics -- Chapter 20. The Population Genetics of Bacteria -- I. Introduction -- II. Genetic Variation and its Interpretation -- III. Species Boundaries and Evolutionary Relationships -- IV. The Importance of Accessory Elements -- V. Experimental Evolution -- VI. The Planned Release of Novel Organisms -- References -- Index.

Note: Front Cover -- Natural Product Biosynthesis by Microorganisms and Plants, Part B -- Copyright -- Contents -- Contributors -- Preface -- Methods in Enzymology -- Section 1: Peptides -- Chapter 1: In Vivo Production of Thiopeptide Variants -- 1. Introduction -- 2. Generation of Thiopeptide Variants -- 2.1. Thiostrepton analogs -- 2.1.1. Overview -- 2.1.2. Thiostrepton variants obtained by precursor peptide mutagenesis -- 2.1.2.1. Deletion of tsrA from the S. laurentii chromosome -- 2.1.2.2. Fosmid engineering to generate complementation vector int-3A10 -- 2.1.2.3. Generation of an intermediate fosmid int-3A100 -- 2.1.2.4. Site-directed mutagenesis of tsrA -- 2.1.2.6. Evaluation of thiostrepton analog production in S. laurentii -- 2.1.3. Gene inactivation to produce thiostrepton analogs -- 2.1.3.1. Deletion of tsrU and tsrT from S. laurentii -- 2.1.3.2. Evaluation of thiostrepton analog production in S. laurentii -- 2.2. Thiocillin analogs -- 2.2.1. Overview -- 2.2.2. Thiocillin analogs obtained by precursor peptide mutagenesis -- 2.2.2.1. Construction of B. cereus DeltatclE-H -- 2.2.2.2. Generation of complementation vector pMGA-tclE-KI -- 2.2.2.3. Evaluation of thiocillin analog production in B. cereus -- 2.3. Nosiheptide analogs -- 2.3.1. Overview -- 2.3.2. Gene inactivation to produce nosiheptide analogs -- 2.3.2.1. Deletion of nosN and nosA in S. actuosus -- 2.3.2.3. Evaluation of nosiheptide analog production in S. actuosus -- References -- Chapter 2: Microviridin Biosynthesis -- 1. Introduction -- 2. Reconstitution In Vitro of the Cyclization Reactions of the Microviridin Biosynthetic Pathway from P. agardhii -- 2.1. Expression of GRASP-like ligase genes mvdC and mvdD -- 2.2. Expression of mvdB -- 2.3. Expression of His6-mvdE -- 2.4. LCMS analysis -- 2.5. Investigation of the lactonization/lactamization reaction in vitro. , 3. Influence of Leader Peptide on the Cyclization Reactions -- 4. Summary -- Acknowledgments -- References -- Chapter 3: Cyclotide Isolation and Characterization -- 1. Introduction -- 2. Isolation of Cyclotides from Plant Tissues -- 3. Isolation of Nucleic Acids Encoding Cyclotide Precursors -- 3.1. Isolating RNA -- 3.2. Primer design for PCR amplification of cyclotide-encoding genes -- 4. Mass Spectrometric Detection and Characterization of Cyclotides -- 4.1. Detection by reduction and alkylation -- 4.2. Sequencing following reduction and digestion -- 5. Structural Analysis of Cyclotides -- 6. Membranolytic Assays of Cyclotides -- 6.1. Preparation of lipid vesicles -- 6.2. Membrane binding by SPR -- 6.3. Vesicle leakage studies -- 7. Summary -- Acknowledgments -- References -- Chapter 4: Ribosomally Encoded Cyclic Peptide Toxins from Mushrooms -- 1. Introduction -- 2. Detection and Purification of Amanita Cyclic Peptide Toxins -- 2.1. Purification of Amanita cyclic peptides -- 2.1.1. Purification from fruiting bodies (mushrooms) -- 2.1.2. Purification from liquid culture of G. marginata -- 2.1.3. Purification of α-amanitin from G. marginata cultured on petri plates -- 3. Identification of Cyclic Peptide Genes in Amanita and Other Fungi -- 4. Purification and Assay of Prolyl Oligopeptidase (POP) from Mushrooms -- 4.1. Isolation of POP from fruiting bodies of C. apala -- 4.1.1. Preparation of crude extract -- 4.1.2. HPLC purification of POP -- 4.1.2.1. Hydrophobic interaction chromatography -- 4.1.2.2. Hydroxyapatite chromatography -- 4.1.2.3. Anion exchange chromatography -- 4.2. Assays for POP -- 4.2.1. Assay with chromogenic substrate -- 4.2.2. Assay with synthetic peptides -- 5. Immunodetection of POP in Mushroom Tissues -- 5.1. Preparation of antibodies -- 5.2. Immunological and microscopic methods -- 6. Summary -- Acknowledgments -- References. , Chapter 5: The Pictet-Spengler Mechanism Involved in the Biosynthesis of Tetrahydroisoquinoline Antitumor Antibiotics: A Novel Function for a Nonribosomal Peptide Synthetase -- 1. Introduction -- 2. Biosynthetic Analysis of THIQ Core Scaffold Assembly -- 2.1. Previous biosynthetic studies on saframycin family members -- 2.2. Biogenesis of saframycin A based on bioinformatic analysis -- 3. Reaction Mechanism for Constructing a Pentacyclic THIQ Core Skeleton Catalyzed by a Novel Function for a Single NRPS Modul -- 4. Comparison Between Typical Peptide Synthesis and the Pictet-Spengler Reaction Catalyzed by NRPS Machinery -- 5. Reaction Profiles Affected by the Chain Length of the Acyl Moiety -- 6. Multiple Roles for the Fatty Acyl Chain During the Maturation Process in Late-Stage Biosynthesis -- 7. Methods for Substrate Synthesis, Enzyme Preparation, and Assay Conditions -- 7.1. Synthesis of the substrates -- 7.1.1. Synthesis of the fatty acyl-peptidyl CoA esters -- 7.1.2. Synthesis of the fatty acyl-peptidyl aldehydes -- 7.2. Overexpression, purification, and phosphopantetheinylation of SfmC -- 7.2.1. Construction of SfmC expression vectors -- 7.2.2. Overexpression and purification -- 7.2.3. Phosphopantetheinylation of SfmC -- 7.3. Enzymatic reactions with saframycin NRPS SfmC -- 7.3.1. SfmC-catalyzed reaction using the fatty-acyl-peptidyl-CoA thioester or the fatty acyl-peptidyl aldehyde -- 7.3.2. Large-scale enzymatic reaction for preparation of presaframycins -- 8. Conclusion -- Acknowledgments -- References -- Section 2: Other Compounds -- Chapter 6: Discovery and Biosynthesis of Phosphonate and Phosphinate Natural Products -- 1. Introduction -- 2. Spectrum of Phosphonate Natural Products: Structures and Bioactivities -- 3. Biosynthetic Pathways -- 4. Screening Bacteria for Phosphonate Producing Gene Clusters. , 4.1. Protocol to identify pepM or ppD genes -- 5. Purification of Phosphonates/Phosphinates -- 5.1. Screening by NMR spectroscopy -- 5.2. Protocol to distinguish between phosphonates and cyclic phosphate esters -- 5.3. Phosphonate purification strategies -- 5.4. Protocol to remove salts -- 6. Structural Elucidation of Phosphonates -- 6.1. 31P NMR spectroscopy -- 6.2. Liquid chromatography-Fourier transform mass spectrometry -- 7. A Wealth of Unusual Biosynthetic Enzymes -- 8. Conclusion and Outlook -- Acknowledgments -- References -- Chapter 7: RlmN and AtsB as Models for the Overproduction and Characterization of Radical SAM Proteins -- 1. Introduction -- 2. RlmN as a Model for Gene Expression and Overproduction of RS Proteins -- 2.1. General cloning strategy -- 2.2. Gene expression and protein overproduction -- 2.3. Protocol for expression of the rlmN gene -- 3. RlmN as a Model for Purification of RS Proteins -- 3.1. Excluding O2 -- 3.2. Protocol for purifying RlmN -- 3.3. Chemical reconstitution of as-isolated RlmN -- 3.4. Protocol for chemical reconstitution of as-isolated RlmN -- 3.5. Removing adventitiously bound iron and sulfide from chemically reconstituted RS proteins -- 4. Overproduction and Purification of E. coli Flavodoxin and Flavodoxin Reductase -- 4.1. Protocol for overexpressing E. coli genes for Flv and Flx -- 4.2. Protocol for purification of Flv or Flx -- 5. AtsB as a Model for Determination of Configuration and Stoichiometry of Iron-Sulfur Clusters in RS Enzymes -- 5.1. Strategy for determining iron-sulfur cluster configuration and stoichiometry -- 5.2. Protocol for preparation of iron-sulfur proteins for amino acid analysis -- 5.3. Protocol for preparation of samples for Mössbauer and EPR spectroscopies -- 6. Activity Determination of RS Enzymes -- 6.1. Protocol for analysis of SAM-related products in RS reactions. , 6.2. Protocol for standard curve preparation of SAM-related products -- 6.3. Protocol for HPLC quantification of SAM-related products -- Acknowledgments -- References -- Chapter 8: Fe(II)-Dependent, Uridine-5-Monophosphate α-Ketoglutarate Dioxygenases in the Synthesis of 5-Modified Nucleosides -- 1. Introduction -- 2. Methods -- 2.1. Bioinformatic analysis of LipL and homologous dioxygenases -- 2.2. Cloning and heterologous expression -- 2.3. SEC-RI-MALLS-ICP-MS to determine metalloprotein stoichiometry -- 2.4. Activity assays -- 2.4.1. Detection of succinate using an enzyme-coupled reaction -- 2.4.2. HPLC analysis -- 2.4.3. Malachite-green binding assay -- 3. Summary -- Acknowledgments -- References -- Section 3: Tailoring Reactions -- Chapter 9: Tailoring Reactions Catalyzed by Heme-Dependent Enzymes: Spectroscopic Characterization of the L-Tryptophan-Nitrating Cytochrom P450 TxtE -- 1. Introduction -- 1.1. Cytochromes P450 -- 1.2. The catalytic mechanisms of CYPs -- 1.3. CYP tailoring enzymes in thaxtomin phytotoxin biosynthesis -- 2. Overexpression, Purification, and Characterization of the Novel Nitrating CYP TxtE -- 2.1. His6-TxtE purification -- 3. Protein Characterization Using UV-vis Spectroscopy -- 3.1. Recording the spectrum of the Fe(III) resting state -- 3.2. Reduction of the ferric heme to ferrous heme using sodium dithionite -- 3.3. Recording the soret spectrum of the ferrous-CO complex -- 4. Ligand and Substrate Binding -- 4.1. Qualitative binding assays: Type I difference spectra -- 4.1.1. Dual beam spectrophotometer -- 4.1.2. Single beam spectrophotometer -- 4.2. Qualitative binding assays: Type II spectra -- 4.2.1. Generation of amine Fe(III) complexes (tryptamine, imidazole, clotrimazole, bifonazole) -- 4.2.2. Generation of an Fe(III)-NO complex -- 5. Summary -- Acknowledgment -- References. , Chapter 10: Oxidative Tailoring Reactions Catalyzed by Nonheme Iron-Dependent Enzymes: Streptorubin B Biosynthesis as an Example.

Note: Cover -- Contents -- Contents by Subject Area -- Preface -- From the Preface to the First Edition -- Guide to the Encyclopedia -- Acknowledgments -- Part A -- Chapter 1. ABC Transport -- Chapter 2. Acetic Acid Production -- Chapter 3. Acetogenesis and Acetogenic Bacteria -- Chapter 4. Actinomycetes -- Chapter 5. Adhesion, Bacterial -- Chapter 6. Aerobic Respiration: Oxidases and Globins -- Chapter 7. Aerosol Infections -- Chapter 8. Agrobacterium -- Chapter 9. Agrobacterium and Plant Cell Transformation -- Chapter 10. AIDS, Historical -- Chapter 11. Airborne Microorganisms and Indoor Air Quality -- Chapter 12. Alkaline Environments -- Chapter 13. Amino Acid Function and Synthesis -- Chapter 14. Amino Acid Production -- Chapter 15. Aminoglycosides, Bioactive Bacterial Metabolites -- Chapter 16. Amylases, Microbial -- Chapter 17. Anaerobic Respiration -- Chapter 18. Antibiotic Biosynthesis -- Chapter 19. Antibodies and B Cells -- Chapter 20. Antifungal Agents -- Chapter 21. Antigenic Variation -- Chapter 22. Antisense RNAs -- Chapter 23. Antiviral Agents -- Chapter 24. Arboviruses -- Chapter 25. Archaea -- Chapter 26. Arsenic -- Chapter 27. Attenuation, Transcriptional -- Chapter 28. Autotrophic CO2 Metabolism -- Chapter 29. Azotobacter -- Part B -- Chapter 1. Bacillus subtilis, Genetics -- Chapter 2. Bacteriocins -- Chapter 3. Bacteriophages -- Chapter 4. Beer/Brewing -- Chapter 5. Beet Necrotic Yellow Vein Virus -- Chapter 6. Biocatalysis for Synthesis of Chiral Pharmaceutical Intermediates -- Chapter 7. Biocides -- Chapter 8. Biodegradation -- Chapter 9. Biodeterioration: In Wood, Architecture, Art, and Other Media -- Chapter 10. Biofilms and Biofouling -- Chapter 11. Biological Control of Weeds -- Chapter 12. Biological Nitrogen Fixation -- Chapter 13. Biological Warfare -- Chapter 14. Bioluminescence, Microbial. , Chapter 15. Biomonitors of Environmental Contamination by Microorganisms -- Chapter 16. Biopesticides, Microbial -- Chapter 17. Biopolymers, Production and Uses of -- Chapter 18. Bioreactor Monitoring and Control -- Chapter 19. Bioreactors -- Chapter 20. Bioremediation -- Chapter 21. Biosensors -- Chapter 22. Biosurfactants -- Chapter 23. Biotransformations -- Part C -- Chapter 1. Carbohydrate Synthesis and Metabolism -- Chapter 2. Carbon and Nitrogen Assimilation, Regulation of -- Chapter 3. Careers in Microbiology -- Chapter 4. Caulobacter, Genetics -- Chapter 5. Cell Division, Prokaryotes -- Chapter 6. Cell Membrane: Structure and Function -- Chapter 7. Cellular Immunity -- Chapter 8. Cellulases -- Chapter 9. Cell Walls, Bacterial -- Chapter 10. Chemotaxis -- Chapter 11. Chlamydia -- Chapter 12. Cholera -- Chapter 13. Cholera, Historical -- Chapter 14. Chromosome, Bacterial -- Chapter 15. Chromosome Replication and Segregation -- Chapter 16. Clostridia -- Chapter 17. Coenzyme and Prosthetic Group Biosynthesis -- Chapter 18. Conjugation, Bacterial -- Chapter 19. Conservation of Cultural Heritage -- Chapter 20. Continuous Culture -- Chapter 21. Cosmetic Microbiology -- Chapter 22. Crystalline Bacterial Cell Surface Layers -- Chapter 23. Cyanobacteria.

Note: Front Cover -- Complex Enzymes in Microbial Natural Product Biosynthesis, Part A: Overview Articles and Peptides -- Copyright -- Contents -- Contributors -- Preface -- Methods in Enzymology -- Chapter 1: Approaches to Discovering Novel Antibacterial and Antifungal Agents -- 1. Introduction -- 2. Strains and Samples -- 3. Targets and Assays for Antibacterial and Antifungal Programs -- 4. Screening -- 5. Hit Follow-up -- 6. Databases, Operations, and Costs -- 7. Perspectives -- Acknowledgment -- References -- Chapter 2: From Microbial Products to Novel Drugs that Target a Multitude of Disease Indications -- 1. Microbial Diversity and Biotechnological Products -- 2. Secondary Metabolites -- 3. Conclusions -- Acknowledgments -- References -- Chapter 3: Discovering Natural Products from Myxobacteria with Emphasis on Rare Producer Strains in Combination with Improved Analytical Methods -- 1. Introduction -- 2. The Search for Novel Myxobacteria and Their Metabolites-Basic Considerations -- 3. Methods for Isolation, Purification, and Preservation of Novel Myxobacteria -- 4. Fermentation and Screening for Known and Novel Metabolites -- Acknowledgment -- References -- Chapter 4: Analyzing the Regulation of Antibiotic Production in Streptomycetes -- 1. Introduction -- 2. The Regulation of Antibiotic Production in Streptomycetes -- 3. Identifying Regulatory Genes for Antibiotic Biosynthesis -- 4. Characterizing Regulatory Genes for Antibiotic Biosynthesis -- Acknowledgments -- References -- Chapter 5: Applying the Genetics of Secondary Metabolism in Model Actinomycetes to the Discovery of New Antibiotics -- 1. Introduction -- 2. Actinomycetes as Antibiotic Factories -- 3. Effects of Culture Conditions and Metabolism -- 4. Molecular Genetic Factors that Regulate Antibiotic Production -- 5. Applications for New Antibiotic Screening Technologies. , 6. Future Prospects -- Acknowledgments -- References -- Chapter 6: Regulation of Antibiotic Production by Bacterial Hormones -- 1. Introduction -- 2. Rapid Small-Scale gamma-Butyrolactone Purification -- 3. Antibiotic Bioassay -- 4. Kanamycin Bioassay -- 5. Identification of gamma-Butyrolactone Receptors -- 6. Identification of the gamma-Butyrolactone Receptor Targets -- 7. Gel Retardation Assay to Detect Target Sequences of the gamma-Butyrolactone Receptors -- 8. Conclusions -- Acknowledgments -- References -- Chapter 7: Cloning and Analysis of Natural Product Pathways -- 1. Introduction -- 2. Cloning and Identification of Biosynthetic Gene Clusters -- 3. Analysis of Natural Product Pathways by PCR-Targeted Gene Replacement -- 4. In Vitro Transposon Mutagenesis -- 5. Heterologous Expression of Biosynthetic Gene Clusters -- 6. Reassembling Entire Gene Clusters by "Stitching" Overlapping Cosmid Clones -- 7. Conclusions -- Acknowledgments -- References -- Chapter 8: Methods for In Silico Prediction of Microbial Polyketide and Nonribosomal Peptide Biosynthetic Pathways from DNA Sequence Data -- 1. Introduction -- 2. Converting Type I PKSs to Structural Elements -- 3. Converting NRPS Domain Strings to Structural Elements -- 4. Concluding Remarks -- Acknowledgments -- References -- Chapter 9: Synthetic Probes for Polyketide and Nonribosomal Peptide Biosynthetic Enzymes -- 1. Introduction -- 2. Synthetic Probes of PKS and NRPS Mechanism -- 3. Synthetic Probes of PKS and NRPS Structure -- 4. Synthetic Probes for Proteomic Identification of PKS and NRPS Enzymes -- 5. Conclusions -- References -- Chapter 10: Using Phosphopantetheinyl Transferases for Enzyme Posttranslational Activation, Site Specific Protein Labeling and Identification of Natural Product Biosynthetic Gene Clusters from Bacterial Genomes -- 1. Introduction -- 2. Experimental Procedures. , 3. Conclusion -- References -- Chapter 11: Sugar Biosynthesis and Modification -- 1. Introduction -- 2. Deoxysugar Biosynthesis -- 3. Deoxysugar Transfer -- 4. Modification of the Glycosylation Pattern through Gene Inactivation -- 5. Modification of the Glycosylation Pattern through Heterologous Gene Expression -- 6. Modification of the Glycosylation Pattern through Combinatorial Biosynthesis -- 7. Gene Cassette Plasmids for Deoxysugar Biosynthesis -- 8. Generation of Glycosylated Compounds -- 9. Tailoring Modifications of the Attached Deoxysugars -- 10. Detection of Glycosylated Compounds -- Acknowledgments -- References -- Chapter 12: The Power of Glycosyltransferases to Generate Bioactive Natural Compounds -- 1. Introduction -- 2. Application of GTs in Producing Unnatural Bioactive Molecules -- References -- Chapter 13: Nonribosomal Peptide Synthetases: Mechanistic and Structural Aspects of Essential Domains -- 1. Introduction -- 2. Mechanistic and Structural Aspects of Essential NRPS Domains -- 3. Structural Insights into an Entire Termination Module -- References -- Chapter 14: Biosynthesis of Nonribosomal Peptide Precursors -- 1. Introduction -- 2. Precursors from Amino Acid Metabolism -- 3. Fatty Acid Precursor Biosynthesis -- 4. Polyketide Precursors -- 5. Glycosyl Building Blocks -- 6. Conclusion -- References -- Chapter 15: Plasmid-Borne Gene Cluster Assemblage and Heterologous Biosynthesis of Nonribosomal Peptides in Escherichia coli -- 1. Introduction -- 2. Biosynthetic Pathway of Nonribosomal Peptides -- 3. Echinomycin Biosynthetic Pathway -- 4. Construction of A Multigene Assembly on Expression Vectors -- 5. Heterologous Gene Expression and NRP Biosynthesis in E. coli -- 6. Self-Resistance Mechanism -- 7. Stability of Transformants Carrying Multiple Very Large Plasmids. , 8. Engineering of Heterologous NRP Biosynthetic Pathways in E. coli -- 9. Conclusion -- References -- Chapter 16: Enzymology of beta-Lactam Compounds with Cephem Structure Produced by Actinomycete -- 1. Introduction -- 2. Biosynthesis of Cephamycins: Enzymes and Genes -- 3. Early Steps Specific for Cephamycin Biosynthesis -- 4. Common Steps in Cephamycin-Producing Actinomycetes and Penicillin- or Cephalosporin-Producing Filamentous Fungi -- 5. Specific Steps for Tailoring the Cephem Nucleus in Actinomycetes -- 6. Regulation of Cephamycin C Production -- Acknowledgments -- References -- Chapter 17: Siderophore Biosynthesis: A Substrate Specificity Assay for Nonribosomal Peptide Synthetase-Independent Siderophore Synthetases Involving Trapping of Acyl-Adenylate Intermediates with Hydroxylamine -- 1. Introduction -- 2. NRPS-Dependent Pathways for Siderophore Biosynthesis -- 3. NRPS-Independent Pathway for Siderophore Biosynthesis -- 4. Hybrid NRPS/NIS Pathway for Petrobactin Biosynthesis -- 5. Hydroxamate-Formation Assay for NIS Synthetases -- References -- Chapter 18: Molecular Genetic Approaches to Analyze Glycopeptide Biosynthesis -- 1. Structural Classification of Glycopeptide Antibiotics -- 2. Methods for Analyzing Glycopeptide Biosynthesis -- 3. Investigation of Glycopeptide Biosynthetic Steps -- 4. Regulation, Self-Resistance, and Excretion -- 5. Linking Primary and Secondary Metabolism -- 6. Approaches for the Generation of New Glycopeptides -- Acknowledgments -- References -- Chapter 19: In Vitro Studies of Phenol Coupling Enzymes Involved in Vancomycin Biosynthesis -- 1. Introduction -- 2. Peptide Synthesis -- 3. Peptide Thioesters -- 4. In Vitro Assays with OxyB -- 5. Production and Purification of Enzymes -- References -- Chapter 20: Biosynthesis and Genetic Engineering of Lipopeptides in Streptomyces roseosporus -- 1. Introduction. , 2. Biosynthesis and Genetic Engineering of Daptomycin in S. rosesoporus -- 3. Sources of Genes for Combinatorial Biosynthesis -- 4. Genetic Engineering of Novel Lipopeptides -- 5. Concluding Remarks -- Acknowledgments -- References -- Chapter 21: In Vitro Studies of Lantibiotic Biosynthesis -- 1. Introduction -- 2. Mining Microbial Genomes for Novel Lantibiotics -- 3. Expression and Purification of Lantibiotic Precursor Peptides (LanAs) -- 4. Expression, Purification, and Assay of LanM Enzymes -- 5. Expression, Purification, and Assays of LanC Cyclases -- 6. The Protease Domain of Class II Lantibiotic Transporters -- 7. Additional Posttranslational Modifications in Lantibiotics -- References -- Chapter 22: Whole-Cell Generation of Lantibiotic Variants -- 1. Introduction -- 2. Variant Generation -- 3. Conclusions -- References -- Chapter 23: Cyanobactin Ribosomally Synthesized Peptides-A Case of Deep Metagenome Mining -- 1. Introduction -- 2. Some Remaining Questions -- 3. Obtaining Prochloron Cells and DNA -- 4. Chemical Analysis -- 5. Cyanobactin Gene Cloning and Identification -- 6. Heterologous Expression in E. coli -- 7. Deep Metagenome Mining -- 8. Enzymatic Analysis of Cyanobactin Biosynthesis -- 9. Applying Deep Metagenome Mining: Pathway Engineering -- Acknowledgments -- References -- Author Index -- Subject Index -- Colour Plates.

Note: Front Cover -- Genetics of Bacterial Diversity -- Copyright Page -- Table of Contents -- Contributors -- Preface -- Section I: Introductory Chapters-the Diversity of Bacteria and of Bacterial Genetics -- Chapter 1. Bacterial Diversity: the Range of Interesting Things that Bacteria Do -- I. INTRODUCTION -- II. ECOLOGICAL NICHES -- III. TOWARDS A PHYLOGENY OF BACTERIA -- IV. REINVENTION THROUGHOUT THE PHYLOGENETIC TREE -- V. SOME UNEXPECTED ATTRIBUTES OF BACTERIA -- V. CONCLUSION -- Acknowledgements -- References -- Chapter 2. Diversity of Bacterial Genetics -- I. INTRODUCTION -- II. THE PROKARYOTIC GENOME -- III. TRANSFER OF CHROMOSOMAL DNA BETWEEN BACTERIA -- IV. GENE EXPRESSION -- V. GENE-, PATHWAY- AND REGULON-SPECIFIC REGULATORY MECHANISMS -- VI. DIFFERENCES BETWEEN PROKARYOTE AND EUKARYOTE GENETICS -- VII. CLOSING REMARKS -- Acknowledgements -- References -- Chapter 3. Cloning and Molecular Analysis of Bacterial Genes -- I. INTRODUCTION -- II. CLONING BACTERIAL DNA -- III. MUTAGENESIS WITH CLONED DNA -- IV. BIOCHEMICAL PROCEDURES THAT EXPLOIT CLONED DNA -- V. CURRENT LIMITATIONS AND POSSIBILITIES -- References -- Section II: Specialized Metabolic Capabilities of Bacteria -- Chapter 4. Regulation of Luminescence in Marine Bacteria -- I. INTRODUCTION -- II. ORGANIZATION AND FUNCTION OF lux GENES -- III. REGULATION OF lux EXPRESSION -- IV. LUMINESCENCE VARIATION -- V. CONCLUSIONS -- Acknowledgements -- References -- Chapter 5. Photosynthesis in Rhodospirillaceae -- I. INTRODUCTION -- II. STRUCTURE-FUNCTION OF THE PHOTOSYNTHETIC APPARATUS -- III. PROTEIN COMPONENTS OF THE PHOTOSYNTHETIC APPARATUS -- IV. PHOTOSYNTHETIC APPARATUS GENES -- V. IN VITRO MUT AGENESIS STUDIES -- VI. GENETIC ENGINEERING IN REACTION CENTRES -- References -- Chapter 6. The Genetics of Nitrogen Fixation -- I. THE DIVERSITY OF NITROGEN-FIXING BACTERIA. , II. THE nif GENES OF KLEBSIELLA PNEUMONIAE -- III. THE ASSEMBLY OF ACTIVE NITROGENASE -- IV. THE BIOCHEMISTRY AND PHYSIOLOGY OF NITROGENASE -- V. THE THREE NITROGENASES OF AZOTOBACTER -- VI. nif GENES IN OTHER ORGANISMS -- VII. REARRANGEMENT OF nif GENES IN ANABAENA -- VIII. REGULATION OF EXPRESSION OF nif GENES -- IX. CONCLUDING REMARKS -- Acknowledgements -- References -- Chapter 7. Antibiotic Biosynthesis in Streptomyces -- I. INTRODUCTION TO STREPTOMYCES BIOLOGY -- II. ANTIBIOTIC PRODUCTION -- III. MOLECULAR GENETICS OF ANTIBIOTIC PRODUCTION -- IV. OVERVIEW, IMPLICATIONS AND PROSPECTS -- References -- Chapter 8. Catabolism of Aromatic Hydrocarbons by Pseudomonas -- I. INTRODUCTION -- II. BIOCHEMICAL STRATEGIES FOR OXIDATIVE CATABOLISM OF AROMATICS -- III. ORGANIZATION AND REGULATION OF GENES FOR CATABOLISM OF AROMATIC HYDROCARBONS -- IV. UTILITY OF DETERMINANTS OF CATABOLIC PATHWAYS -- V. LABORATORY EVOLUTION OF AROMATIC CATABOLIC PATHWAYS -- VI. CONCLUDING REMARKS -- Acknowledgements -- References -- Chapter 9. Mercury Resistance in Bacteria -- I. INTRODUCTION -- II. BACTERIAL TRANSFORMATIONS OF MERCURY -- III. MERCURY RESISTANCE GENES -- IV. THE GRAM-NEGATIVE STRUCTURAL GENES AND THEIR PRODUCTS -- V. A MODEL FOR MERCURY RESISTANCE IN BACTERIA -- VI. REGULATION OF EXPRESSION OF THE MERCURY RESISTANCE GENES -- VII. OVERVIEW AND PROSPECTS -- Acknowledgements -- References -- Section III: Morphologica lDifferentiation-Flagella, Spores and Multicellular Development -- Chapter 10. Differentiation in Caulobacter: Flagellum Development, Motility and Chemotaxis -- I. INTRODUCTION -- IL DEVELOPMENTAL PROGRAMMES AND CELL DIFFERENTIATION -- III. REGULATION OF FLAGELLUM BIOSYNTHESIS -- IV. CONTROL OF CHEMOTAXIS AND POSITIONING OF DIFFERENTIATED STRUCTURES -- V. PROSPECTS-THE CELL CYCLE AS A REGULATOR OF TEMPORAL AND SPATIAL PATTERNING -- Acknowledgements. , References -- Chapter 11. Pathways of Developmentally Regulated Gene Expression in Bacillus subtilis -- I. INTRODUCTION -- II. SPORULATION AND GERMINATION -- III. GENES INVOLVED IN SPORULATION AND GERMINATION -- IV. DEVELOPMENTAL GENES ARE SWITCHED ON IN AN ORDERED TEMPORAL SEQUENCE -- V. COMPARTMENTALIZATION OF GENE EXPRESSION -- VI. DEPENDENCE PATTERNS OF DEVELOPMENTAL GENE EXPRESSION: FOUR EXAMPLES -- VII. PATHWAYS OF DEVELOPMENTALLY REGULATED GENE EXPRESSION -- VIII. OVERVIEW, IMPLICATIONS AND PROSPECTS -- Acknowledgements -- References -- Chapter 12. Multiceïlular Development in Myxobacteria -- I. INTRODUCTION -- II. FRUITING BODY DEVELOPMENT FOLLOWS A PROGRAMME -- III. OPERON FUSIONS EXPOSE A PROGRAMME OF DIFFERENTIAL GENE EXPRESSION -- IV. CELL INTERACTIONS COORDINATE THE PROGRAMME OF FRUITING BODY DEVELOPMENT -- V. MUTANTS OF GROUPS A, B, C AND D DIFFER GENETICALLY -- VI. EXPRESSION OF β-GALACTOSIDASE FROM lac FUSION STRAINS DEPENDS ON THE PRODUCTS OF THE asg, bsg, csg AND dsg GENES -- VII. A-FACTOR AND C-FACTOR ACTIVITIES CAN BE FOUND IN CELL EXTRACTS -- VIII. THE asg, bsg, csg AND dsg GENE LOCI CAN BE ISOLATED -- IX. OVERVIEW AND PROSPECTS -- References -- Section IV: Bacterial Adaptations to Animal Pathogenicity -- Chapter 13. The Molecular Basis of Antigenic Variation in Pathogenic Neisseria -- I. INTRODUCTION -- II. DIVERSITY AND VIRULENCE -- III. GENETIC MECHANISMS FOR PILUS VARIATION -- IV. GENETIC MECHANISMS FOR P.II VARIATION -- V. CONCLUSIONS -- Acknowledgements -- References -- Chapter 14. Adhesins of Pathogenic Escherichia coli -- I. INTRODUCTION -- II. BACTERIAL ADHERENCE TO ANIMAL TISSUES -- III. ADHESIN GENETICS -- IV. EVOLUTIONARY PERSPECTIVES -- References -- Chapter 15. Genetic Studies of Enterotoxin and Other Potential Virulence Factors of Vibriocholerae -- I. INTRODUCTION -- II. CHOLERA TOXIN GENES. , III. ADHERENCE AND COLONIZATION -- IV. OTHER POTENTIAL VIRULENCE FACTORS -- V. REGULATION OF VIRULENCE GENE EXPRESSION -- VI. PERSPECTIVES -- References -- Chapter 16. Iron Scavenging in the Pathogenesis of Escherichia coli -- I. INTRODUCTION -- II. ENTEROBACTERIAL IRON UPTAKE SYSTEMS -- III. MOLECULAR GENETICS -- IV. BIOCHEMICAL GENETICS -- V. REGULATION -- VI. EPILOGUE -- References -- Section V: Bacteria that Interact with Plants as Parasites or Symbionts -- Chapter 17. Pathogenicity of Xanthomonas and Related Bacteria Towards Plants -- I. INTRODUCTION -- II. STRATEGIES AND TECHNIQUES FOR STUDYING THE GENETICS OF PATHOGENICITY -- III. FUNCTION OF SOME PATHOGENICITY GENES -- IV. CONCLUDING REMARKS -- References -- Chapter 18. Tumorigenicity of Agrobacterium on Plants -- I. INTRODUCTION -- II. Ti AND Ri PLASMIDS -- III. T-DNA -- IV. GENES AND SEQUENCES NECESSARY FOR T-DNA TRANSFER -- V. DIFFERENT STEPS IN THE PROCESS OF TUMOUR INDUCTION -- VI. PROSPECTS FOR APPLICATIONS -- References -- Chapter 19. The Symbiosis Between Rhizobium and Legumes -- I. INTRODUCTION -- II. METHODS FOR IDENTIFYING BACTERIAL GENES INVOLVED IN NODULATION -- III. POLYSACCHARIDE SYNTHESIS IS IMPORTANT FOR NODULATION -- IV. ANALYSIS OF NOD GENE FUNCTION -- V. REGULATION OF NOD GENE TRANSCRIPTION -- VI. CONCLUSIONS -- References -- Section VI: Bacterial Population Genetics -- Chapter 20. The Population Genetics of Bacteria -- I. INTRODUCTION -- II. GENETIC VARIATION AND ITS INTERPRETATION -- III. SPECIES BOUNDARIES AND EVOLUTIONARY RELATIONSHIPS -- IV. THE IMPORTANCE OF ACCESSORY ELEMENTS -- V. EXPERIMENTAL EVOLUTION -- VI. THE PLANNED RELEASE OF NOVEL ORGANISMS -- References -- Index.

Note: Literaturverz. S. 229 - 240 , Actinomycetes and antibiotics -- Antibiotic discovery and resistance -- Microbial sex -- Towards gene cloning -- From chromosome map to DNA sequence -- Bacteria that develop -- The switch to antibiotic production -- Unnatural natural products -- Functional genomics -- Genomics against tuberculosis and leprosy