USE OF SYNCHROTRON X-RAY FLUORESCENCE MICROTOMOGRAPHY TO CHARACTERIZE THE GENES

使用同步加速器 X 射线荧光显微术来表征基因

基本信息

  • 批准号:
    8362435
  • 负责人:
  • 金额:
    $ 0.33万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2011
  • 资助国家:
    美国
  • 起止时间:
    2011-03-01 至 2012-02-29
  • 项目状态:
    已结题

项目摘要

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Our research combines established genomic techniques, such as gene deletion and overexpression, with synchrotron x-ray fluorescence (SXRF) microspectroscopy and microtomography to characterize the function of genes involved in metal ion homeostasis in plants. We pioneered the use of SXRF in gene characterization with a gene critical for vacuolar iron (Fe) transport in seed, which has far reaching implications for the development of Fe enriched grain and the reduction in Fe deficiency anemia in humans. While mature seed represents an ideal sample for SXRF analysis, seed-expressed genes are only a subset of the plant genome. To characterize genes that encode for metal(loid) transporters in roots, leaves and developing seed, fresh or live plant imaging is preferred, due to the elemental and distributional artifacts that can result from dehydration, resin infiltration and sectioning. Constraints on live or fresh plant imaging range from the sensitivity of the detector, the motion of the stepping motors, as well as a range of issues with sample drying. We have conducted preliminary studies at BL2-3 that have allowed us to collect tomograms of Fe and other important elements from the excised stem of a 2 day old Arabidopsis seedling in approximately one hour, analysis that would have taken 10 hours at other environmental microprobe beamlines. By comparing the wild type tomogram with that of a mutant lacking a vacuolar iron transporter (VIT1) we have shown clear distributional contrasts that have shed new light on the function of the gene. We wish to conduct fresh- and live-plant imaging studies at BL2-3 to help characterize genes involved in metal ion homeostasis. Our current projects involve further characterizing genes that encode for vacuolar Fe transporter, VIT1, and transporters responsible for the transport of arsenic within the plant and into the seed.
这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 子项目的主要研究者可能是由其他来源提供的, 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量, 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 我们的研究结合了现有的基因组技术,如基因缺失和过表达,同步辐射X射线荧光(SXRF)显微光谱和显微断层扫描,以表征植物中金属离子稳态相关基因的功能。我们率先使用SXRF技术对种子中液泡铁(Fe)转运的关键基因进行基因表征,这对富铁谷物的开发和人类缺铁性贫血的减少具有深远的意义。虽然成熟种子代表了SXRF分析的理想样品,但种子表达的基因只是植物基因组的一个子集。为了表征根、叶和发育中的种子中编码金属(类金属)转运蛋白的基因,优选新鲜或活植物成像,这是由于脱水、树脂渗透和切片可能导致的元素和分布伪影。对活体或新鲜植物成像的限制包括检测器的灵敏度、步进电机的运动以及样品干燥的一系列问题。我们已经在BL 2 -3进行了初步研究,这使我们能够在大约1小时内从2天大的拟南芥幼苗的切除茎中收集Fe和其他重要元素的断层图像,在其他环境微探针光束线处需要10小时的分析。通过比较野生型断层扫描与缺乏液泡铁转运蛋白(VIT 1)的突变体,我们已经显示出明确的分布对比,揭示了新的基因的功能。我们希望在BL 2 -3进行新鲜和活体植物成像研究,以帮助表征参与金属离子稳态的基因。我们目前的项目涉及进一步表征基因编码液泡铁转运蛋白,维生素T1,转运蛋白负责运输砷在植物和种子。

项目成果

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Tracy Punshon其他文献

Tracy Punshon的其他文献

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{{ truncateString('Tracy Punshon', 18)}}的其他基金

SXRF IMAGING OF LIVE ARABIDOPSIS PLANTS TO CHARACTERIZE GENES INVOLVED IN METAL
对活拟南芥植物进行 SXRF 成像,以表征与金属相关的基因
  • 批准号:
    8362267
  • 财政年份:
    2011
  • 资助金额:
    $ 0.33万
  • 项目类别:
SXRF IMAGING OF LIVE ARABIDOPSIS PLANTS TO CHARACTERIZE GENES INVOLVED IN METAL
对活拟南芥植物进行 SXRF 成像,以表征与金属相关的基因
  • 批准号:
    8170259
  • 财政年份:
    2010
  • 资助金额:
    $ 0.33万
  • 项目类别:
Assessing Maternal-Fetal Exposure Pathways Using Bio-Imaging
使用生物成像评估母婴暴露途径
  • 批准号:
    8627626
  • 财政年份:
  • 资助金额:
    $ 0.33万
  • 项目类别:
Assessing Maternal-Fetal Exposure Pathways Using Bio-Imaging
使用生物成像评估母婴暴露途径
  • 批准号:
    8465524
  • 财政年份:
  • 资助金额:
    $ 0.33万
  • 项目类别:

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