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噬菌体肽库筛选隐丹参酮高亲和性结合肽
引用本文:龙军,袁冬平,陆茵,林洁.噬菌体肽库筛选隐丹参酮高亲和性结合肽[J].中国药科大学学报,2013,44(5):465-469.
作者姓名:龙军  袁冬平  陆茵  林洁
作者单位:南京中医药大学药学院;南京中医药大学药理学教研室;南京中医药大学江苏省中药药效与安全性评价重点实验室;云南大学生命科学学院
基金项目:国家自然科学基金资助项目(No.81173174);江苏省自然科学基金资助项目(No.SBK201241602);江苏省高校自然科学基金资助项目(No.11KJB360004)
摘    要:通过噬菌体十二肽库,以隐丹参酮为靶分子,筛选与隐丹参酮具有高亲和性的结合短肽。经过噬菌体十二肽库的3轮淘选,获得阳性噬菌体克隆,挑选结合力强的克隆进行测序,得到隐丹参酮的高亲和性结合短肽序列,并进行生物信息学分析。本试验共筛选到10个隐丹参酮的高亲和性结合短肽序列,有614种蛋白质与其结构相匹配。多肽序列的核心序列为VILDFGEI。本研究获得了与隐丹参酮结合的高亲和性多肽,为深入研究隐丹参酮的作用靶点以及分子作用机制提供了实验依据和结构基础。

关 键 词:隐丹参酮  噬菌体肽库  多肽  生物信息学

Screening of high affinity peptides binding to cryptotanshinone with phage display peptide library
LONG Jun,YUAN Dongping,LU Yin and LIN Jie.Screening of high affinity peptides binding to cryptotanshinone with phage display peptide library[J].Journal of China Pharmaceutical University,2013,44(5):465-469.
Authors:LONG Jun  YUAN Dongping  LU Yin and LIN Jie
Abstract:To screen peptides with high affinity to cryptotanshinone, using cryptotanshinone as a ligand, phage clones binding to cryptotanshinone were screened with phage display 12-mer peptide library through three rounds of biopanning. After binding activity was measured by ELISA, the phage clones displaying high affinity to cryptotanshinone were selected and amplified. DNAs of phage clones were extracted and sequenced, and the coding peptide sequences were analyzed by bioinformatics. Amino acid sequences of 10 positive clones with high affinity to cryptotanshinone were identified. 614 proteins matched partially to the peptide found by homological retrieval with BLAST. The core peptide sequence VILDFGEI was deduced by this study. Peptides with high affinity to cryptotanshinone were obtained through the experiment, which provided experimental evidence and structural foundations for further study of the targets of effects and molecular mechanisms of actions of cryptotanshinone.
Keywords:cryptotanshinone  phage display peptide library  peptide  bioinformatics
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