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关于苯丙氨酸转运RNA诱导荧光寡核苷酸与核糖体解码位点的结合

On the Phe-tRNA induced binding of fluorescent oligonucleotides to the ribosomal decoding site.

作者信息

Menzel H M

出版信息

Nucleic Acids Res. 1977 Aug;4(8):2881-92. doi: 10.1093/nar/4.8.2881.

Abstract

Fluorescent oligonucleotides were prepared by dansylation of 5'-amino uridylates of varying chainlength. Except for the trinucleoside diphosphate, they stimulated the binding of PhetRNA TO 70S E. coli ribosomes as efficiently as underivatised oligouridylic acids of comparable chainlength. The ternary ribosomal complex [70S X Phe-tRNA X dansyl-n5'U(pU)4] was separated from excess oligonucleotide and its fluorescence spectra were measured. The quantum yield of the dansylated pentauridylate was enhanced 2.5 fold when bound to the ribosomal decoding site, but no shift of the emission spectrum was observed. The ribosomal complex is considered useful for topographic investigations by singlet energy transfer, using the functionally defined decoding site as reference point.

摘要

通过对不同链长的5'-氨基尿苷酸进行丹磺酰化反应制备了荧光寡核苷酸。除了三磷酸核苷二磷酸外,它们刺激苯丙氨酰-tRNA与70S大肠杆菌核糖体结合的效率与具有可比链长的未衍生化寡尿苷酸一样高。将三元核糖体复合物[70S×苯丙氨酰-tRNA×丹磺酰化的n5'U(pU)4]与过量的寡核苷酸分离,并测量其荧光光谱。当与核糖体解码位点结合时,丹磺酰化的五聚尿苷酸的量子产率提高了2.5倍,但未观察到发射光谱的位移。核糖体复合物被认为可用于通过单重态能量转移进行拓扑学研究,以功能定义的解码位点作为参考点。

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