Determination of the sub-cellular location of sex pheromone biosynthesis in the yellow mealworm beetle, Tenebrio molitor

ii ACKNOWLEDGMENTS iii TABLE OF CONTENTS iv LIST OF TABLES v LIST OF FIGURES vi 1.0. INTRODUCTION 1 2.0. MATERIALS AND METHODS 8 2.1. Insect Cultures 8 2.2. Purchased Chemicals 8 2.3. Buffers 9 2.4. Synthesis of Required Chemicals 9 2.5. Dissections 11 2.6. Preparation of Homogenate 11 2.7. Subcellular Fractionation 11 2.8. In vitro Enzyme Assays 12 2.9. Extraction, purification and quantification of [3H]-4-methylnonanol 13 2.10. Protein Assay 14 2.11. Marker Enzymes Analysis 14 2.12. Statistical analysis 15 3.0. DATA AND RESULTS 16 3.1. Synthesis of Required Chemical 16 3.2. Biochemical Studies 16 3.2.1. Preliminary In vitro Experiment 16 3.2.2. Aldehyde In vitro Experiments 17 3.2.3. Fatty acyl CoA In Vitro Experiments 17 3.2.4. Protein Curve 18 3.2.5. Marker Enzyme Assays 18 4.0. DISCUSSION 26 4.1. Preliminary Experiments 26 4.2. Isolation of 4-Methylnonanoyl CoA reductase 27 4.3. Further Studies 30 REFERENCES 32