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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___________________________________________________________________________________Chapter 4: Discussion4.1: PCR screen<strong>in</strong>g assays for detect<strong>in</strong>g biosynthetic capability <strong>in</strong> environmentalact<strong>in</strong>obacteria ________________________________________________ 1124.1.1 Design <strong>of</strong> PCR primers _______________________________________ 1134.1.2 Type II Polyketide Synthase ___________________________________ 1134.1.3 Type I Polyketide Synthase ____________________________________ 1154.1.4 dTDP – glucose synthase______________________________________ 1174.1.5 Isopenicill<strong>in</strong> N synthase_______________________________________ 1184.2: Secondary metabolite production <strong>of</strong> act<strong>in</strong>obacteria__________________ 1214.2.1 Solid Agar _________________________________________________ 1214.2.2 Submerged Fermentations _____________________________________ 1214.2.2.1 Carbon Sources __________________________________________ 1214.2.2.2 Nitrogen sources _________________________________________ 1224.2.2.3 Suitability <strong>of</strong> liquid media for secondary metabolite screen<strong>in</strong>g ____ 1234.2.2.4 Effect <strong>of</strong> oil supplementation to submerged fermentations ________ 1234.2.2.5 Duration for chemical expression <strong>of</strong> bioactive metabolites ________ 1244.3: Correlation between genetic screen<strong>in</strong>g <strong>and</strong> antibiotic effects __________ 1244.4: Adaptation <strong>of</strong> cultivation conditions for secondary metaboliteScreen<strong>in</strong>g ____________________________________________________ 1264.4.1 Solid Substrate Fermentations __________________________________ 1264.5: Extractability <strong>of</strong> bioactive metabolites ____________________________ 1274.6: Isolation <strong>of</strong> Bioactive Metabolites _______________________________ 1284.7: UV-Vis Spectroscopy scann<strong>in</strong>g <strong>of</strong> organic extracts__________________ 1294.8: Analysis <strong>of</strong> bioactive organic extracts by Reverse-Phase High PerformanceLiquid Chromatography (RP-HPLC) with UV-Visible Diode Array <strong>and</strong>Electrospray Ionisation- mass Spectrometric (ESI-MS) detection ____ 1304.9: Conclusions __________________________________________________ 132APPENDIX 1: Conference presentation <strong>and</strong> AwardsA1.1: Conference Presentation _____________________________________ 134A1.2: Awards____________________________________________________ 134REFERENCES ___________________________________________________ 135___________________________________________________________________________________
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