Gryaznov S M, Schultz R, Chaturvedi S K, Letsinger R L
Lynx Therapeutics Inc., Hayward, CA 94545.
Nucleic Acids Res. 1994 Jun 25;22(12):2366-9. doi: 10.1093/nar/22.12.2366.
A new approach to increase the selectivity of interaction between oligonucleotide probes and target nucleic acids is described. In place of a single, relatively long oligonucleotide probe, two or three short oligomers terminated by thiophosphoryl and bromoacetamido groups are employed. Fast and efficient autoligation takes place when the oligomers hybridize in a contiguous mode to the same complementary strand such that a thiophosphoryl group on one strand and a bromoacetamido group on another are brought into proximity. A single nucleotide mismatch for the short probes leads to marked reduction in the rate of autoligation. The binding affinity of the product is close to that for a natural probe of the same length. This approach could have potential in oligonucleotide-based diagnostics, chemical amplification systems, and therapeutic applications.
描述了一种提高寡核苷酸探针与靶核酸之间相互作用选择性的新方法。采用的不是单个相对较长的寡核苷酸探针，而是两个或三个由硫代磷酰基和溴乙酰胺基终止的短寡聚物。当寡聚物以连续模式与同一条互补链杂交，使得一条链上的硫代磷酰基和另一条链上的溴乙酰胺基靠近时，会发生快速而有效的自动连接。短探针的单个核苷酸错配会导致自动连接速率显著降低。产物的结合亲和力与相同长度的天然探针相近。这种方法在基于寡核苷酸的诊断、化学扩增系统和治疗应用中可能具有潜力。
