含有2'-脱氧-2'-氟核苷的寡核苷酸探针用于大肠杆菌RNase H对RNA的区域特异性切割。
The use of oligonucleotide probes containing 2'-deoxy-2'-fluoronucleosides for regiospecific cleavage of RNA by RNase H from Escherichia coli.
作者信息
Schmidt S, Niemann A, Krynetskaya N F, Oretskaya T S, Metelev V G, Suchomlinov V V, Shabarova Z A, Cech D
机构信息
Humboldt University, Institute of Bioorganic Chemistry, Berlin, Germany.
出版信息
Biochim Biophys Acta. 1992 Feb 28;1130(1):41-6. doi: 10.1016/0167-4781(92)90459-d.
Protected 2'-deoxy-2'-fluorouridine and 2'-deoxy-2'-fluorocytidine suitable for incorporation into oligonucleotides via the phosphoramidite approach have been prepared. Five modified and two unmodified oligonucleotides have been synthesized to investigate the regiospecific cleavage of a 5S RNA from Escherichia coli by RNase H. In order to show whether the modified oligonucleotides are able to hybridize with the RNA the physico-chemical properties (melting curves, CD spectra) of analogous DNA/oligodeoxyribonucleotide duplexes have been examined. The modified oligonucleotides are shown to form stable duplexes with a DNA-matrix which exist in an A-like form. Two of the modified probes containing four 2'-deoxy-2'-fluorocytidines or two 2'-deoxy-2'-fluorouridines direct the splitting by RNase H of only one phosphodiester bond of the RNA.
已制备出适合通过亚磷酰胺法掺入寡核苷酸的2'-脱氧-2'-氟尿苷和2'-脱氧-2'-氟胞苷。合成了五条修饰的和两条未修饰的寡核苷酸，以研究核糖核酸酶H对大肠杆菌5S RNA的区域特异性切割。为了表明修饰的寡核苷酸是否能够与RNA杂交，已检测了类似DNA/寡脱氧核糖核苷酸双链体的物理化学性质（熔解曲线、圆二色光谱）。结果表明，修饰的寡核苷酸与呈A样形式存在的DNA模板形成稳定的双链体。两条含有四个2'-脱氧-2'-氟胞苷或两个2'-脱氧-2'-氟尿苷的修饰探针可引导核糖核酸酶H仅切割RNA的一个磷酸二酯键。
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