A Toolkit for in Vivo Visualization/Modulation of Plant Cell Wall Polysaccharides

植物细胞壁多糖体内可视化/调节的工具包

• 批准号: 0923992
• 负责人: Michael Hahn
• 金额: $ 434.04万
• 依托单位: University of Georgia Research Foundation Inc
• 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 2017-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0923992&HistoricalAwards=false
• 关键词: Toolkit; Vivo; Visualization; Modulation; Plant

PI: Michael G. Hahn (University of Georgia)CoPIs: Geert-Jan Boons, Harry Gilbert, Christopher S. King, Malcolm A. O'Neill, and William S. York (University of Georgia)There are currently no tools that can be used for in vivo imaging plant cell wall polysaccharides and glycoproteins. This project will provide a set of molecular probes for cell wall polysaccharides in the form of single-chain antibody fragments (ScFvs) and carbohydrate-binding modules (CBMs) as tools for in vivo imaging of plant cell walls using microscopy. The specific goals are: 1) to generate, either from hybridoma cell lines or from carbohydrate binding domains, a comprehensive series of tagged molecular probes that can be expressed in plant cells and will bind to cell wall glycans in vivo; and 2) to demonstrate that the probes generated will permit the localization of diverse carbohydrate structures within living plant cells or alternatively alter cell wall structure by selectively disrupting polysaccharide incorporation into walls in transgenic plants. It is anticipated that the molecular probes generated during the course of this project will be instrumental to those interested in the dynamics of plant cell wall polysaccharides and that some of these probes will disrupt cell wall assembly and structure through their interaction with the polysaccharides/glycoproteins and serve as valuable tools to discover the functions of cell wall polysaccharides in the biology of plant cells. With respect to outreach and training, project personnel will teach a one week course to be held during each summer at the Complex Carbohydrate Research Center during the tenure of the grant to teach polysaccharide localization methods to interested scientists. An explicit effort will be made to involve undergraduate students in the research carried out under this grant. The participation of historically underrepresented groups in the research will be fostered by recruiting undergraduate students through the University of Georgia Summer Undergraduate Research Program (SURP) (http://www.grad.uga.edu/outreach&diversity/surp.html). All students and post-doctoral associates will be closely mentored to encourage and foster their development as research scientists. Information about all antibodies, ScFvs, and CBMs generated through this project, will be made available immediately to the scientific community via posting to a web-based antibody database (http://www.wallmabdb.net). All molecular probes will be made available to the research community without restriction through CarboSource (http://www.carbosource.net). Plasmids and transgenic plant lines generated in the molecular probe function validation studies will be available through the Arabidopsis Biological Resource Center (http://abrc.osu.edu).

PI：Michael G.哈恩（格鲁吉亚大学）CoPIs：Geert-Jan Boons，Harry吉尔伯特，Christopher S.作者：Malcolm A.奥尼尔和威廉S.约克（格鲁吉亚大学）目前还没有可用于植物细胞壁多糖和糖蛋白体内成像的工具。 该项目将提供一套单链抗体片段（ScFvs）和碳水化合物结合模块（CBM）形式的细胞壁多糖分子探针，作为使用显微镜进行植物细胞壁体内成像的工具。 具体目标是：1）从杂交瘤细胞系或从碳水化合物结合结构域产生可在植物细胞中表达并将在体内结合细胞壁聚糖的全面系列的标记分子探针;和2）为了证明所产生的探针将允许在活的植物细胞内定位不同的碳水化合物结构，或者通过选择性地破坏多糖掺入来改变细胞壁结构，在转基因植物的墙壁上。 预计在本项目过程中产生的分子探针将对那些对植物细胞壁多糖动力学感兴趣的人有所帮助，其中一些探针将通过与多糖/糖蛋白的相互作用破坏细胞壁组装和结构，并作为发现植物细胞生物学中细胞壁多糖功能的有价值的工具。 关于推广和培训，项目人员将在赠款有效期内每年夏季在复杂碳水化合物研究中心教授为期一周的课程，向感兴趣的科学家教授多糖定位方法。 将作出明确的努力，让本科生参与这项赠款下进行的研究。 将通过格鲁吉亚大学夏季本科生研究计划（SURP）（http：//www.grad.uga.edu/outreach diversity/surp.html）招募本科生，促进历史上代表性不足的群体参与研究。 所有的学生和博士后助理将密切指导，以鼓励和促进他们的发展作为研究科学家。 关于通过该项目产生的所有抗体、ScFvs和CBM的信息将通过发布到基于网络的抗体数据库（http：//www.example.com）立即提供给科学界。www.wallmabdb.net 所有分子探针将通过CarboSource（http：//www.example.com）无限制地提供给研究团体。www.carbosource.net 分子探针功能验证研究中产生的质粒和转基因植物系可通过拟南芥生物资源中心（http：//www.example.com）获得。abrc.osu.edu