Chlamydomonas reinhardtii Ergosterol Gene Complementation Experiments

麦角甾醇是莱茵衣藻膜中发现的主要甾醇。麦角甾醇是一种最终产物，主要存在于酵母菌等真菌生物中。虽然过去的研究已经发现了一些麦角甾醇突变体，但对该物种的甾醇生物合成途径知之甚少。这种模式生物，莱因哈蒂梭菌，是非常独特的，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允许确定参与麦角甾醇生物合成的潜在基因。有了这些知识，我们设计了一个通路的工作模型。最终，本项目的目标是利用分子生物学技术，用莱茵衣藻麦角甾醇基因补充酵母中的甾醇突变。互补将使我们能够扩增莱茵衣藻的甾醇基因，将这些基因转移到先前表征的酵母，酿酒酵母的甾醇突变体中，并表征其在异源物种中的功能。更好地了解这一途径和所涉及的基因也将为这个物种的进化以及可能的其他藻类系统提供新的信息。此外，它还将促进对该模型系统中脂质生物合成的理解。更广泛的影响更广泛的影响：作为培养最多STEM（科学、技术、工程和数学）学生并继续获得博士学位的国家领导者，泽维尔大学在科学方面一直保持着卓越的声誉。学生将有机会参与一个结合脂质生物化学和分子生物学技术的项目，这些技术将为他们在其他机构的研究生院的严格要求进行培训和准备。该项目将有助于继续支持那些有兴趣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.