RUI: Cloning and Characterization of OPENED IMMATURE FLOWER

智力优势：许多基因表达的研究都集中在转录水平上，而选择性剪接（一种增加转录物多样性的保守机制）相对未被探索。该项目旨在通过确定植物剪接网络中的新因素来加深对选择性剪接的理解。在拟南芥中，SR 45属于在剪接事件中起重要作用的因子家族。基因开放未成熟花（OIF）最近被确定为SR 45的遗传增强子。为了理解OIF如何调节SR 45的剪接调节功能，将通过基于图位的克隆和通过将候选OIF基因转化到对于SR 45和OIF是突变体的植物中的功能互补来分离OIF基因。剪接网络中的新因子将通过筛选在sr 45突变体背景中产生开放的未成熟花表型的额外增强子来寻找。将通过扫描电子显微镜进一步表征突变表型，并使用RT-PCR分析潜在基因靶点的剪接模式。 总之，这些方法将为植物发育中选择性剪接的作用提供新的见解。更广泛的影响：本科生将从事一个独特的研究经验，可以在其他本科院校复制。首先，通过圣文德大学和马里兰州大学之间的伙伴关系，学生将通过在两个机构获得第一手实验室经验和参加国家和国际会议来接触积极的研究。第二，一个新的综合课程，植物发育和生理学，包括基于研究的讲座，开放式的研究实验室会议，和互动讨论，将设计并提供给大二/大三/大四的本科生。在每次实验课之前，将指定一名学生并对其进行培训，以便在讲师的帮助下领导实验。来自全班的学生将参与制作一张海报，描述他们所有实验室课程的综合结果，并分享合著权。将选出一名学生在会议上展示海报。这种创新的设计将使学生进行批判性思维，积极的研究和创造性的讨论。在这个项目中提供给学生的所有机会都旨在激发热情和积累研究经验，为未来的科学研究职业做准备。

Intellectual Merit: Many studies of gene expression have focused on transcription levels, leaving alternative splicing, a conserved mechanism that increases transcript diversity, relatively unexplored. This project seeks to deepen understanding of alternative splicing by identifying new factors within the splicing network of plants. In Arabidopsis thaliana, SR45 belongs to a family of factors that play important roles in splicing events. The gene OPENED IMMATURE FLOWER (OIF) was recently identified as a genetic enhancer of SR45. To understand how OIF modulates the splicing regulatory function of SR45, the OIF gene will be isolated by map-based cloning and functional complementation by transformation of candidate OIF genes into plants that are mutant for SR45 and OIF. New factors in the splicing network will be sought by screening for additional enhancers that produce an opened immature flower phenotype in the sr45 mutant background. Mutant phenotypes will be further characterized by scanning electron microscopy, and the splicing patterns of potential gene targets will be analyzed using RT-PCR. Together, these approaches will provide new insights into the role of alternative splicing in plant development. Broader Impact: Undergraduate students will be engaged in a unique research experience that can be replicated at other undergraduate institutions. First, through a partnership between St. Bonaventure University and the University of Maryland, students will be exposed to active research by gaining first-hand laboratory experience at both institutions and by attending national and international conferences. Second, a new integrative course, Plant Development and Physiology, including research-based lectures, open-ended research lab sessions, and interactive discussions, will be designed and offered to undergraduates at the sophomore/junior/senior level. Prior to each lab session, a student will be designated and trained to lead the lab with help from the instructor. Students from the entire class will participate in generating a poster describing their combined results from all lab sessions and share co-authorship. One student will be selected to present the poster at a conference. This innovative design will engage students in critical thinking, active research and creative discussion. All the opportunities provided to the students in this project are aimed at stimulating enthusiasm and building research experience in preparation for future careers in scientific research.