Chlamydomonas reinhardtii Ergosterol Gene Complementation Experiments

麦角甾醇是莱茵衣藻细胞膜中的主要甾醇。麦角甾醇是一种最终产物，主要存在于酵母等真菌生物体中。虽然过去的研究已经在莱茵梭菌中发现了一些麦角固醇突变体，但对该物种中固醇的生物合成途径知之甚少。模式生物，C.reinhardtii，非常独特，因为它在膜中积累麦角甾醇。随着衣藻基因组的阐明，生物信息学分析已经能够确定与麦角固醇生物合成有关的潜在基因。有了这些知识，已经设计了该通路的工作模型。最终，该项目的目标是使用分子生物学技术来补充来自莱茵衣藻的麦角甾醇基因酵母中的甾醇突变。互补将使我们能够从莱茵衣藻中扩增出甾醇基因，将这些基因转移到酵母的甾醇突变体酿酒酵母中，并鉴定它们在异源物种中的功能。更好地了解这一途径和涉及的基因也将提供关于该物种以及可能其他藻类系统进化的新信息。此外，它还将促进对这一模型系统中脂肪生物合成的理解。布罗德影响：作为培养出最多继续获得博士学位的STEM(科学、技术、工程和数学)学生的全国领先者，泽维尔大学在科学方面享有卓越的声誉。学生将有机会参加一个包含脂质生物化学以及分子生物学技术的项目，这些技术将培训他们并为他们在其他机构研究生院的严酷环境做好准备。该项目将有助于继续支持有志于在科学领域追求成功事业的学生的基础设施。

Ergosterol is the major sterol found in the membranes of Chlamydomonas reinhardtii. Ergosterol is an end-product sterol primarily found in fungal organisms like yeast. While past studies have identified some ergosterol mutants in C. reinhardtii, very little is known about the sterol biosynthesis pathway in this species. The model organism, C. reinhardtii, is very unique in the sense that it accumulates ergosterol in its membranes. With the elucidation of the Chlamydomonas genome, bioinformatics analysis has allowed the determination of potential genes involved in ergosterol biosynthesis. With this knowledge, a working model of the pathway has been designed. Ultimately, the goal of this project is to employ molecular biology techniques to complement sterol mutations in yeasts with ergosterol genes from Chlamydomonas reinhardtii. Complementation would allow us to amplify the sterol genes from Chlamydomonas reinhardtii, transfer those genes into the previously characterized sterol mutants of the yeast, Saccharomyces cerevisiae, and characterize their function in a heterologous species. Better understanding of this pathway and the genes involved will also provide new information about the evolution of this species and possibly other algal systems. In addition, it will advance the understanding of lipid biosynthesis in this model system.Broader ImpactsBroader Impacts: As the national leader in producing the largest number of STEM (Science, Technology, Engineering and Mathematics) students who go on to receive Ph.Ds, Xavier University has upheld a reputation for excellence in the sciences. Students will have an opportunity to participate in a project that incorporates lipid biochemistry, as well as molecular biology techniques that will train and prepare them for the rigors of graduate school at other institutions. This project will help to continue to support the infrastructure of students who are interested in pursuing successful careers in the sciences.