Strategic Reprogramming of the Ergot Alkaloid Pathway

7. 项目总结/文摘

7. Project Summary/Abstract Ergot alkaloids improve human health as powerful and versatile pharmaceuticals for treatment of multiple conditions including senile dementia, Alzheimer’s disease, Parkinson’s disease, migraines, hyperprolactinemia, and type 2 diabetes. Tremendous diversity in structure and activity can be found among the natural and semi-synthetic ergot alkaloids. Small changes in structure may lead to a great changes in activity. The long-term goal of this research program is to understand and ultimately control the biosynthesis of diverse ergot alkaloids by determining the functions of genes and gene products that produce and diversify ergot alkaloids. The requested equipment, a Waters Arc HPLC (high performance liquid chromatography) system, will replace an aging instrument that presents frequent maintenance and data management challenges. The new instrument will enhance efficiency of the research team and improve the quality of data. Working with the new instrument will provide undergraduate and graduate students with relevant training. The instrument will be used to gather data for each of the approaches being used for both specific aims of the parent award, titled “Strategic reprogramming of the ergot alkaloid pathway.” Both specific aims involve modifying fungal ergot alkaloid pathways by CRISPR/Cas9-based approaches or expression of heterologous genes, and then analyzing the altered ergot alkaloid profiles by HPLC and subsequent mass spectrometry approaches. The new HPLC will have an integral role in gathering data for all of the planned experimental approaches. The HPLC will be critical in identifying which candidate strains have been mutated by the recombinant DNA approaches and in identifying the metabolites accumulating in those constructed strains. The HPLC also will be used to collect quantitative data on ergot alkaloid accumulation which will be used in statistical analyses to test hypotheses about gene function. Results of the proposed project will reveal roles of specific genes and provide strains of fungi that produce molecules with pharmaceutical significance. Additional benefits include meaningful experiences for graduate and undergraduate students and further development of platforms for modification and improvement of additional or novel ergot alkaloids.