Nucleotide Modification and Polymerase Engineering for Creating a Novel Class of Artificial Nucleic Acid Aptamers

摘要: Enzymatic transcription and reverse transcrip- tion of artificial nucleic acids would be an important technique to allow the application random screening methods such as system- atic evolution ligands by exponential enrichment (SELEX), Non-SELEX selection, one-step selection etc. These able identify various functional aptamers ribozymes with functions similar antibodies enzymes, which could useful not only research tools for molecular biology but also diag- nostic agents, therapeutic drugs Recently, KOD its re- lated DNA polymerases have been used preparing modified acids, including base-modified nu- cleic sugar-modified ones phosphate- ones. However, thus far, reasons effectiveness polymerase purposes clearly elucidated. Therefore, using mutated poly- merases, we studied here their catalytic properties upon enzy- matic incorporation nucleotide analogues base/sugar/ phosphate modifications. Experimental data indicate that characteristic kinetic enabled recognition nucleotides. Among those mutants, one achieved efficient successive bridged nucleo- tides a 2�c -ONHCH2CH2-4�c linkage, prom- ising candidates acid drugs. In this study, char- acteristic nucleoside triphosphates were shown, effec- tiveness genetic engineering in improvement enzyme polymerization has demon- strated.