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首页> 外文学位 >Part I: Spectroscopic characterization of sol-gel-derived materials and dendritic compounds amenable to optical chemical sensor development. Part II: Steps towards the development of general aminoacylation of transfer RNA molecules via RNA-based catalysis: On-column synthesis and subsequent purification in a multiple turnover system.
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Part I: Spectroscopic characterization of sol-gel-derived materials and dendritic compounds amenable to optical chemical sensor development. Part II: Steps towards the development of general aminoacylation of transfer RNA molecules via RNA-based catalysis: On-column synthesis and subsequent purification in a multiple turnover system.

机译：第一部分：溶胶-凝胶衍生的材料和适合光学化学传感器开发的树枝状化合物的光谱表征。第二部分：通过基于RNA的催化作用，对转移RNA分子进行一般氨基酰化的步骤：柱上合成以及随后在多周转系统中的纯化。

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摘要

Sol-gel-processed materials, which are made under room temperature conditions, can accommodate a variety of dopant molecules within their silica matrix, including thermally labile chemical and biochemical recognition elements. Moreover, under carefully chosen condition of hydrolysis, condensation, and curing, one can tune the morphology and internal dimensions of the resulting glass or ceramic, making these materials ideally suited for chemical/biochemical sensor applications. Described herein, Part I of this thesis investigates the response of environmentally sensitive fluorescent probes, trapped within the silica ‘cage’ of sol-gel-derived materials, in order to provide both qualitative and quantitative description of the microenvironments present within. Specifically, we use steady state and time-resolved fluorescent techniques to measure the solvent polarity, accessibility, and molecular dynamics surrounding the dopant molecule.; Part II of this thesis provides an introduction into the realm of RNA-based catalysis, and the notion of an RNA world. In addition to being a provocative theory capable of shedding light on primitive life during early earth, the study of RNA catalysis holds great promise as a readily accessible and adaptable catalytic system with a wide range of tunable substrate recognition elements. Using in vitro selection, our laboratory has more recently devised an aminoacyl transfer catalyst, or ribozyme (r24mini), which is capable of charging the 3′ terminus of transfer RNA (tRNA) with an aminoacyl ester moiety. Aminoacyl-tRNA is of critical importance during modern-day translation of proteins and is responsible for the maintenance of fidelity between the genome and corresponding phenotype. However, similarly charged tRNA containing nonnatural or analog amino acids also offer a valuable route in the in vitro synthesis of tailor made proteins. Part II of this thesis describes recent efforts undertaken in our laboratory to perform such aminoacylation on-column, in a multiple turnover catalytic system. Moreover, we show that such on-column aminoacylation facilitates the rapid chromatographic separation and purification of aminoacylated-tRNA. Using a variety of bioconjugative methods, we have since immobilized r24mini to stationary agarose columns, in order to perform on-column aminoacylation and purify the resulting ammoacyl-tRNA. The details of each of these methods, and the corresponding results in percent aminoacylated-tRNA are described within.
机译：溶胶-凝胶处理的材料在室温条件下制成，可以在其二氧化硅基体中容纳各种掺杂剂分子，包括热不稳定的化学和生物化学识别元素。此外，在精心选择的水解，冷凝和固化条件下，人们可以调节所得玻璃或陶瓷的形态和内部尺寸，使这些材料非常适合化学/生化传感器应用。本文所述，本论文的第一部分研究了对环境敏感的荧光探针的响应，该探针被困在溶胶-凝胶衍生材料的二氧化硅“笼子”中，以提供对其中存在的微环境的定性和定量描述。具体来说，我们使用稳态和时间分辨荧光技术来测量溶剂极性，可及性以及掺杂剂分子周围的分子动力学。本文的第二部分介绍了基于RNA的催化领域以及RNA世界的概念。 RNA催化研究除了是能够在早期地球上阐明原始生命的挑衅性理论外，还具有广阔的前景，因为它具有易于使用的 和 adaptable 催化系统。广泛的可调基板识别元件。通过体外选择，我们的实验室最近设计了一种氨基酰基转移催化剂或核酶（r24mini），该催化剂能够在转移RNA（tRNA）的3 末端带有氨基酰基酯部分。氨酰基-tRNA在现代蛋白质翻译过程中至关重要，并负责维持基因组和相应表型之间的保真度。然而，带有类似电荷的tRNA包含非天然或类似氨基酸也为定制蛋白质的体外合成提供了一条有价值的途径。本文的第二部分描述了在我们的实验室中最近为在多周转催化体系中进行这种氨基酰化的努力。此外，我们表明这种柱上的氨酰化促进了氨基酰化tRNA的快速色谱分离和纯化。自那时以来，我们已使用多种生物偶联方法将r24mini固定在固定的琼脂糖柱上，以进行柱上氨基酰化并纯化所得的氨酰基tRNA。每种方法的详细信息以及相应的氨基酰化tRNA百分比结果均在其中描述。

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  • 作者

    Bonzagni, Neil J.;

  • 作者单位

    State University of New York at Buffalo.;

  • 授予单位 State University of New York at Buffalo.;
  • 学科 Chemistry Analytical.
  • 学位 Ph.D.
  • 年度 2002
  • 页码 279 p.
  • 总页数 279
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 化学;
  • 关键词

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