Facchini, PeterGroves, Ryan A2016-01-142016-01-142016-01-142016Groves, R. A. (2016). Transcriptome Profiling in Ephedra sinica and Catha edulis Reveals Enzymes Putatively Involved in Ephedrine Alkaloid Biosynthesis (Master's thesis, University of Calgary, Calgary, Canada). Retrieved from https://prism.ucalgary.ca. doi:10.11575/PRISM/28507http://hdl.handle.net/11023/2747Amphetamine analogs are a class of medicinal compounds produced by Ephedra sinica and Catha edulis. Two commonly used members of this class of compounds include the cough suppressant (1S-2S)-pseudoephedrine, and the stimulant (1R-2S)-ephedrine. These secondary metabolites are produced in planta through a L-phenylalanine derived multi-step pathway. Despite the importance of this biosynthetic pathway, only one enzyme in the pathway has been characterized at the molecular level. Recently, two new transcriptomes have been created for E. sinica and C. edulis with the hopes of facilitating future gene discovery in this untapped pathway. In this investigation, I outlined an efficient transcriptomics based approach to potential gene candidate selection for all putative stages of the amphetamine analog biosynthetic pathway. This investigation also describes a complete approach to molecular cloning, protein expression, and assaying of gene candidates, resulting in the discovery of a key biosynthetic enzyme involved in amphetamine analog biosynthesis in E. sinica.engUniversity of Calgary graduate students retain copyright ownership and moral rights for their thesis. You may use this material in any way that is permitted by the Copyright Act or through licensing that has been assigned to the document. For uses that are not allowable under copyright legislation or licensing, you are required to seek permission.BioinformaticsGeneticsMicrobiologyBiology--MolecularBiochemistryChemistry--PharmaceuticalEphedraKhatAlkaloidsBiosynthesisDatabaseEnzymesRNA sequencingBenzaldehydesEphedrineAmphetamine-analogueTranscriptomemaster thesis10.11575/PRISM/28507