Request fund to purchase equipment (the BioComp gradient primer) to supplement R15 GM139107-01

申请资金购买设备（BioComp梯度引物）以补充R15 GM139107-01

基本信息

批准号：
10582006
负责人：
Ke Zhang Reid
金额：
$ 3.93万
依托单位：
WAKE FOREST UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2024-08-31
项目状态：
已结题

项目摘要

Project Summary (Abstract): We are applying for a supplement fund to purchase a BioComp gradient primer for fractionating density gradients. This equipment is essential for the success of the parent award as several proposed experiments, including polysome profiling, ribosome profiling, and Translating Ribosome Affinity Purification (TRAP), rely on this equipment to receive consistent and reproducible results. We desperately need this equipment to continue our investigation following equipment malfunction of our current system that cannot be repaired (system age and non-availability of parts). Additionally, no other laboratory at our institution has similar equipment we could share. Our talented undergraduate and graduate students will use this equipment to address an exciting but disputed question: does the functional specialization of ribosomes exist to support selective translation? We focus on two paralogs of predicted large ribosomal subunit 27 (Rpl27) found in fission yeast. Although these paralogs have 90% amino acid similarity and are highly conserved through much of eukaryotic evolution, their mutants demonstrate distinct phenotypes. Our preliminary data suggest that they might be part of heterogeneous ribosomes that regulate the translation of different mRNA transcripts. Dissecting the functional differences between Rpl27 paralogs may provide valuable insight into how ribosome protein paralogs potentially play a role in the heterogeneity of ribosomes, inducing a conformational change that ultimately results in altered activity and preference of mRNA transcripts accessible for translation.

项目摘要（摘要）：我们正在申请补充基金，以购买生物制作梯度底漆分级密度梯度。该设备对于父母的成功至关重要作为几个建议的实验奖，包括多核体分析，核糖体分析和翻译核糖体亲和力净化（陷阱），依靠此设备获得一致且可重现的结果。我们迫切需要这种设备在我们当前系统的设备故障之后，继续我们的调查无法修复（零件的系统年龄和不可用）。另外，没有其他我们机构的实验室有类似的设备，我们可以分享。我们的才华本科生和研究生将使用此设备来解决令人兴奋但有争议的问题：是否存在核糖体的功能专业化来支持选择性翻译？我们专注于两个预测的大核糖体亚基27 （RPL27）在裂变酵母中发现。尽管这些旁系同源物具有90％的氨基酸相似性并通过大部分真核进化而高度保守，它们的突变体展示不同的表型。我们的初步数据表明它们可能是调节不同mRNA转录物的翻译的异质核糖体的。剖析RPL27旁系同源物之间的功能差异可能会提供有价值的深入了解核糖体蛋白旁系同源物如何在异质性中发挥作用核糖体，诱导构象变化，最终导致活性改变和mRNA转录本可访问的偏好。