PROBING RIBOSOMAL FUNCTION

化学合成的DNA寡聚体长度小于10个碱基， 用于原位探测核糖体RNA不同区域的功能。 DNA寡聚体将被合成以便与某些DNA寡聚体互补。 rRNA的暴露区域，与这些区域杂交，测定特异性 结合，然后测定复合物以确定功能是否 由于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可能的。 通过使用RNase H剪切异源双链体区域中的rRNA来确保， 然后对所得片段进行短距离测序， 从滑的方向。 将使用的功能测定包括 蛋白质合成，tRNA结合到P位点和A位点， 起始复合物形成和mRNA结合测定。 预计在 完整核糖体中rRNA特定区域的离散功能将 以这种方式决定。

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.