CATALYTIC SITE-SPECIFIC CLEAVAGE OF A DNA-TARGET BY AN OLIGONUCLEOTIDE CARRYING BLEOMYCIN A5

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.