Influence of DNA aptamer structure on the specificity of binding to Taq DNA polymerase

被引:20
|
作者
Yakimovich, OY
Alekseev, YI
Maksimenko, AV
Voronina, OL
Lunin, VG
机构
[1] All Russia Inst Agr Biotechnol, Moscow 127550, Russia
[2] JSC Syntol, Moscow 127550, Russia
[3] Russian Acad Sci, Bach Inst Biochem, Moscow 117871, Russia
关键词
DNA ligands; DNA aptamer design; Taq DNA polymerase; DNA-protein complex; PCR;
D O I
10.1023/A:1022609714768
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The secondary structure of DNA aptamer to Taq DNA polymerase was established as a hairpin. Both stem and loop structures of DNA ligand were shown to be involved in the interaction with Taq DNA polymerase. Moreover, the structure and sequence of DNA aptamer that was the most effective inhibitor of DNA polymerase activity were established. This crucial structure was evaluated as a GC-rich stem longer than 17 bp, and a loop consisting of 12 bases with strictly determined nucleotide sequence. It was demonstrated that nucleotide in position 23 counting from the 5'-end of DNA ligand was involved in direct contact with Taq DNA polymerase. The ability of optimized DNA aptamer TQ21-11 to form a complex with the enzyme was increased 5-fold in comparison to the initial aptamer.
引用
收藏
页码:228 / 235
页数:8
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