通过指数富集的配体系统进化技术筛选抑制人乙酰胆碱酯酶的RNA分子。
Screening of RNA molecules inhibiting human acetylcholinesterase by virtue of systematic evolution of ligands by exponential enrichment.
作者信息
Zhang Xing-Mei, Shao Ning-Sheng, Chi Mu-Gen, Sun Man-Ji
机构信息
Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences, Beijing 100850, China.
出版信息
Acta Pharmacol Sin. 2003 Jul;24(7):711-4.
AIM
To acquire the specific RNA aptamers inhibiting human red blood cell (RBC) acetylcholinesterase (AChE).
METHODS
Systematic evolution of ligands by exponential enrichment (SELEX) aptamer against human red blood cell membrane AChE was selected by microtiter plate method in vitro. The specifity binding to AChE was determined by gel mobility shift analysis. Microcolorispectrophotometric method was used to measure the activity of AChE.
RESULTS
The aptamers to human RBC AChE were identified by 9 reiterative rounds. At the same concentration (2.25 micromol/L), the aptamers did not bind to the recombinant human butyrylcholinesterase (rhBChE) but specifically bound to human RBC AChE and inhibited the enzyme activity.
CONCLUSION
It is an effective way to isolate the specific AChE inhibitor from the vast oligonucleotide combinatorial library by virtue of SELEX.
目的
获得抑制人红细胞（RBC）乙酰胆碱酯酶（AChE）的特异性RNA适配体。
方法
采用微孔板法体外筛选针对人红细胞膜AChE的指数富集配体系统进化（SELEX）适配体。通过凝胶迁移率变动分析确定与AChE的特异性结合。采用微量比色分光光度法测定AChE的活性。
结果
经过9轮重复筛选，鉴定出了人红细胞AChE的适配体。在相同浓度（2.25微摩尔/升）下，该适配体不与重组人丁酰胆碱酯酶（rhBChE）结合，但能特异性结合人红细胞AChE并抑制其酶活性。
结论
借助SELEX从庞大的寡核苷酸组合文库中分离特异性AChE抑制剂是一种有效的方法。
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