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Catalytic site-specific cleavage of a DNA-target by an oligonucleotide carrying bleomycin A5.

机译:带有博来霉素A5的寡核苷酸对DNA靶的催化位点特异性切割。

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摘要

Oligonucleotide reagents have been created which are capable of catalytic site-specific cleavage of DNA-targets. The oligonucleotide reagent Blm-R-pd(CCAAACA) bearing the bleomycin A5 (Blm-RH) residue was used to degrade the DNA-target pd(TGTTTGGCGAAGGA). It has been shown that at equimolar reagent: target concentration the bleomycin oligonucleotide derivative can repeatedly cleave the target at G9, G7, T5, T4 and T3 in site-specific manner. This paper demonstrates that with a 10-fold excess of the DNA-target relative to the reagent 30% degradation of the target was observed primarily at a single position G7. The paper also shows that one reagent molecule containing bleomycin A5 residue was capable to degrade three molecules of the DNA-target. The catalytic activity of Blm-R-pd(CCAAACA) was the highest in the temperature range close to the melting temperature of the reagent-target complex, that is under conditions where the oligonucleotide reagent can form a complementary complex and easily dissociate to interact with the next molecule of the target. The number of target molecules degraded by the bleomycin reagent is limited by the degradation of the antibiotic residue itself.
机译:已经创建了能够催化DNA靶位点特异性切割的寡核苷酸试剂。带有博来霉素A5(Blm-RH)残基的寡核苷酸试剂Blm-R-pd(CCAAACA)用于降解DNA靶标pd(TGTTTGGCGAAGGA)。已经显示在等摩尔试剂:靶标浓度下,博来霉素寡核苷酸衍生物可以以位点特异性方式在G9,G7,T5,T4和T3处重复切割靶标。本文证明,相对于试剂,DNA靶标过量10倍时,主要在单个位置G7上观察到靶标的30%降解。该论文还表明,一个含有博来霉素A5残基的试剂分子能够降解三个DNA靶分子。 Blm-R-pd(CCAAACA)的催化活性在接近试剂-靶标复合物的解链温度的温度范围内最高,即在寡核苷酸试剂可以形成互补复合物且易于解离以与之相互作用的条件下目标的下一个分子。被博来霉素试剂降解的靶分子的数量受到抗生素残基自身降解的限制。

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