Pieken W A, Olsen D B, Aurup H, Williams D M, Heidenreich O, Benseler F, Eckstein F
Max-Planck-Institut für Experimentelle Medizin, Göttingen, FRG.
Nucleic Acids Symp Ser. 1991(24):51-3.
Hammerhead ribozymes containing 2'-fluoro- or 2'-aminonucleotides were prepared by automated chemical synthesis. Incorporation of 2'-fluorouridines, 2'-fluorocytidines or 2'-aminouridines did not appreciably decrease catalytic activity. The presence of 2'-aminocytidines, however, reduced the activity about 20-fold. No catalytic activity could be measured for ribozymes in which all adenosines were replaced by the 2'-fluoro analogue in presence of MgCl2. No single position could be found responsible for this loss of activity. In an attempt to construct ribozymes to hydrolyse HIV-RNA in the 5'-LTR region several constructs were tested on synthetic substrate as well as on run-off transcripts of about 1000 nucleotides length.
通过自动化学合成制备了含有2'-氟代或2'-氨基核苷酸的锤头状核酶。掺入2'-氟尿苷、2'-氟胞苷或2'-氨基尿苷并没有明显降低催化活性。然而,2'-氨基胞苷的存在使活性降低了约20倍。在MgCl2存在的情况下,所有腺苷都被2'-氟类似物取代的核酶无法检测到催化活性。没有发现单一位置对此活性丧失负责。为了构建在5'-LTR区域水解HIV-RNA的核酶,对几种构建体在合成底物以及约1000个核苷酸长度的径流转录本上进行了测试。