PROBING RIBOSOMAL FUNCTION

化学合成的长度小于10个碱基的DNA低聚物将被 用于原位检测核糖体RNA各区域的功能。 将合成DNA寡聚体，以便与某些特定的 RRNA暴露区域，与这些区域杂交，检测其特异性 结合，然后将测定复合体，以确定是否功能 核糖体的活性因DNA探针的存在而受损。 互补DNA与特定rRNA位点的特异性结合将是 通过使用RNaseH来剪裁异源双链区的rRNA来确保 然后对所产生的片段进行短距离测序 从滑行开始的方向。将使用的功能分析将包括 蛋白质合成，结合到P位和A位的tRNA， 起始复合体形成和信使核糖核酸结合试验。预计 完整核糖体中rRNA特定区域的离散功能 以这样的方式来决定。

Chemically-synthesized DNA oligomers less than ten bases long are to be used to probe the functions of various regions of ribosomal RNA in situ. The DNA oligomers will be synthesized so as to be complementary to certain exposed regions of rRNA, hybridized to those regions, assayed for specific binding and then the complex will be assayed to ascertain if the function of the ribosome has been impaired by the presence of the DNA probe. Specific binding of the complementary DNA to a specific rRNA site will be assured by using RNase H to clip the rRNA in the heteroduplex region and then sequencing the resulting fragments for short distances in either direction from the slip. The functional assays to be used will include protein synthesis, tRNA binding to both the P site and the A site, initiation complex formation and mRNA binding assays. It is expected that discrete functions of specific regions of rRNA in the intact ribosome will be determined in this manner.