Inoue H, Hayase Y, Iwai S, Ohtsuka E
Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan.
Nucleic Acids Symp Ser. 1987(18):221-4.
To cleave RNA molecules using RNase H in a site-specific manner, a short deoxyoligonucleotide (3-5mer) joining with 2'-O-methyl oligonucleotide(s) was designed as a DNA splint to be used. Model experiments were carried out using ribooligonucleotide substrates (9 and 18 mer). It was found that the use of this type of splints (9 mer) causes a unique cleavage by RNase H. For example, when 3'm (GA)d(AGAA)m(GGU)5' was used as a hybridization strand, 32pUCUUUCUUCUUCCAGGAU was cleaved specifically between U11 and C12 to yield 32pUCUUUCUUCUU. This method will have a variety of applications for the study of RNA.
为了以位点特异性方式使用核糖核酸酶H切割RNA分子，设计了一种与2'-O-甲基寡核苷酸连接的短脱氧寡核苷酸（3 - 5聚体）作为DNA夹板来使用。使用核糖寡核苷酸底物（9聚体和18聚体）进行了模型实验。发现使用这种类型的夹板（9聚体）会导致核糖核酸酶H进行独特的切割。例如，当使用3'm(GA)d(AGAA)m(GGU)5'作为杂交链时，32pUCUUUCUUCUUCCAGGAU在U11和C12之间被特异性切割，产生32pUCUUUCUUCUU。该方法在RNA研究中将有多种应用。
