PROBING RIBOSOMAL FUNCTION

这项研究的首要目的是阐明 核糖体。具体地说，暴露的、单链的功能 将对核糖体RNA的区域进行分析，以确定它们如何影响 核糖体的功能。通过这种方式，tRNA、MPNA和 可以确定平移系数绑定。此外， RNA的结构作为核糖体在翻译中的功能可以 观察到的。尤其令人感兴趣的是抗生素结合的部位， 由于核糖体功能或结构在这些疾病中受到明显影响 案子。这些实验的设计是用化学方法合成短片。 与核糖体RNA特定区域互补的DNA寡聚体， 将这些DNA探针与核糖体上的靶点杂交，并检测 从而对tRNA结合、mRNA结合、翻译因子产生影响 绑定或抗生素绑定。相对结合将通过以下方法进行检测 监测放射性标记探针的残留结合活性 或使用硝酸纤维素滤膜分析的配体。场地的专用性 将使用RNaseH确定绑定。使用这些结果，它应该 有可能开始了解mRNA、tRNA、翻译因子是如何 核糖体从事合成蛋白质的业务。

The overriding purpose of this research is to elucidate the function of the ribosome. Specifically the function of exposed, single-stranded regions of ribosomal RNA will be assayed to determine how they affect the function of the ribosome. In this way the sites where TRNA, MPNA and translation factors bind can be determined. In addition, changes in the structure of the RNA as the ribosome functions in translation can be observed. Of particular interest are the sites where antibiotics bind, since ribosomal function or structure is markedly affected in these cases. The design of the experiments is to chemically synthesize short DNA oligomers complementary to specific regions of ribosomal RNA, hybridize these DNA probes to target sites on the ribosome, and assay the resulting affects on tRNA binding, mRNA binding, translation factor binding or antibiotic binding. Relative binding will be assayed by monitoring the residual binding activity of radioactively-labeled probes or ligands using nitrocellulose filter assays. Specificity of site binding will be determined using RNase H. Using these results it should be possible to begin to understand how mRNA, tRNA, translation factors and the ribosome go about the business of synthesizing proteins.