Detection and Expression of Biosynthetic Genes in Actinobacteria ...
Detection and Expression of Biosynthetic Genes in Actinobacteria ...
Detection and Expression of Biosynthetic Genes in Actinobacteria ...
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___________________________________________________________________________________1.4.1.2 Codon Usage (CU) __________________________________________27Section 5: Genetic Factors Affect<strong>in</strong>g the Production <strong>of</strong> SecondaryMetabolites ______________________________________________271.5.1 Plasmid Instability ____________________________________________271.5.2 Induced Mutagensis ___________________________________________27Section 6: Molecular Techniques used to Detect SecondaryMetabolites <strong>Biosynthetic</strong> <strong>Genes</strong> _____________________________281.6.1 DNA Homology <strong>of</strong> Secondary Metabolite <strong>Biosynthetic</strong> <strong>Genes</strong> (SMBG)<strong>in</strong> Act<strong>in</strong>obacteria______________________________________________291.6.2 <strong>Detection</strong> <strong>of</strong> Secondary Metabolite <strong>Biosynthetic</strong> <strong>Genes</strong> <strong>in</strong> Act<strong>in</strong>obacteriaus<strong>in</strong>g Heterologous DNA Probes _________________________________291.6.3 <strong>Detection</strong> <strong>of</strong> Secondary Metabolite <strong>Biosynthetic</strong> <strong>Genes</strong> us<strong>in</strong>g PolymeraseCha<strong>in</strong> Reaction _______________________________________________321.6.3.1 PCR Screen<strong>in</strong>g <strong>of</strong> SMBG <strong>in</strong> Natural Act<strong>in</strong>obacterial Populations ______331.6.3.2 PCR Clon<strong>in</strong>g <strong>and</strong> Screen<strong>in</strong>g <strong>of</strong> SMBG <strong>in</strong> Act<strong>in</strong>obacteria_____________341.6.4 Access<strong>in</strong>g Secondary Metabolite Diversity from UnculturedSoil Microorganisms___________________________________________35Section 7: Production <strong>of</strong> Secondary Metabolites ________________________361.7.1 Physical Conditions Affect<strong>in</strong>g Secondary Metabolite Production ________371.7.2 Submerged Fermentation (SmF) Conditions Affect<strong>in</strong>g SecondaryMetabolite Production__________________________________________371.7.2.1 Components <strong>of</strong> fermentation media_____________________________381.7.2.2 Macronutrients _____________________________________________381.7.2.2.1 Carbon Source __________________________________________381.7.2.2.2 Nitrogen Source _________________________________________391.7.2.2.3 Phosphate Source________________________________________391.7.2.2.4 Sulfur, Potassium, Magnesium Sources_______________________401.7.2.3 Inorganic <strong>and</strong> Organic Micronutrients __________________________411.7.2.4 Temperature Shifts _________________________________________421.7.2.5 pH Effects _______________________________________________441.7.2.6 Dissolved Oxygen__________________________________________441.7.2.7 Precursors, C<strong>of</strong>actors <strong>and</strong> Nucleotides __________________________451.7.2.8 Enzyme Inhibitors <strong>and</strong> Repressors <strong>of</strong> Secondary Metabolism _______451.7.2.9 Autoregulators_____________________________________________461.7.2.10 Miscellaneous Factors Influenc<strong>in</strong>g Secondary MetaboliteProduction________________________________________________461.7.3 Solid State Fermentations _____________________________________47Section 8: Genetic Modification <strong>of</strong> <strong>Biosynthetic</strong> <strong>Genes</strong> <strong>and</strong> the Development <strong>of</strong>Novel Compounds _________________________________________481.8.1 Genetic manipulation <strong>of</strong> secondary metabolite produc<strong>in</strong>gAct<strong>in</strong>obacteria________________________________________________481.8.2 Metabolic Eng<strong>in</strong>eer<strong>in</strong>g <strong>of</strong> Secondary metabolite Produc<strong>in</strong>gAct<strong>in</strong>obacteria________________________________________________49___________________________________________________________________________________
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