RUI: Cloning and Characterization of OPENED IMMATURE FLOWER

RUI：开放未成熟花的克隆和表征

• 批准号: 0950158
• 负责人: Xiao-Ning Zhang
• 金额: $ 21.83万
• 依托单位: Saint Bonaventure University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2015-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0950158&HistoricalAwards=false
• 关键词: RUI; Cloning; Characterization; OPENED; IMMATURE

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.

智力优势：许多关于基因表达的研究都集中在转录水平上，而对选择性剪接--一种增加转录多样性的保守机制--的研究相对较少。这个项目试图通过确定植物剪接网络中的新因素来加深对替代剪接的理解。在拟南芥中，SR45属于一个在剪接事件中发挥重要作用的因子家族。基因开放的未成熟花(OIF)最近被鉴定为SR45的遗传增强子。为了了解OIF如何调控SR45的剪接调控功能，我们将通过基于图谱的克隆和将候选OIF基因转化到SR45和OIF突变的植物中进行功能互补来分离OIF基因。将通过筛选在SR45突变背景中产生开放的未成熟花表型的额外增强剂来寻找剪接网络中的新因素。突变表型将进一步通过扫描电子显微镜进行表征，潜在基因靶标的剪接模式将使用RT-PCR进行分析。总之，这些方法将为替代剪接在植物发育中的作用提供新的见解。更广泛的影响：本科生将参与一种独特的研究体验，这种体验可以在其他本科院校复制。首先，通过圣博纳文迪亚大学和马里兰大学之间的伙伴关系，学生将通过在这两个机构获得第一手实验室经验并参加国内和国际会议，获得积极的研究机会。第二，将设计一门新的综合课程《植物发育与生理学》，包括以研究为基础的讲座、开放式研究实验室课程和互动讨论，并将提供给大二/初三/高年级的本科生。在每次实验之前，将指定一名学生并对其进行培训，让其在教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，描述他们所有实验课程的综合结果，并分享共同撰写的成果。将选出一名学生在一次会议上展示海报。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是为了激发学生的热情和积累研究经验，为未来的科学研究职业生涯做准备。