从噬菌体随机十二肽库中筛选NF-κB(p65)亲和肽的研究  

Screening of phage-displayed random peptide library for binding peptide of NF-κB(p65)

作　　者：闫海涛[1] 马晓芸[1] 董泗建[1] 郑建全[1] 丁日高[1] 

机构地区：[1]军事医学科学院毒物药物研究所,北京100850

出　　处：《军事医学科学院院刊》2005年第6期523-524,527,共3页Bulletin of the Academy of Military Medical Sciences

基　　金：全军十五指令课题(01L054)

摘　　要：目的:采用噬菌体展示技术筛选核转录因子-κB(nuc lear factor-κB,NF-κB)亚基p65亲和肽。方法:以p65亚基为靶分子,利用噬菌体随机十二肽库进行亲和筛选,用ELISA法进行阳性克隆结合特异性验证。结果:经过3轮亲和筛选后,随机挑选15个噬菌体克隆,对其亲和力做ELISA试验,其中6个克隆显示较强的阳性结果。将上述6个阳性克隆DNA测序,序列均为CGTCAGCCTCGGCGAATAAGTATCCGAAGGAGTAAA,相应的氨基酸序列为FTPSDTYSPRLT。竞争性ELISA的结果显示,抑制率随p65亚基浓度的增大而升高。结论:从噬菌体随机肽库中筛选到能与NF-κB p65亚基特异性结合的克隆,并确定了其核心序列。Objective：To screen the p65 subunit of N F-κB binding peptides by phage-display technique. Methods：The p65 subunit of NF-κB was targeted for screening of a random bacteriophage 12-mers peptide library, and the positive clones were identified with ELISA. Results： After three rounds of screening, 6 of 15 phage clones were identified as positive by sandwish ELISA. All of these 6 clones had same DNA sequences： CGTCAGCCTCGGCGAATAAGTATCCGAAGGAGTAAA, and their corresponding peptide sequences were FTPSDTYSPRLT. Such positive clone could bind well to p65 subunit of NF-κB , and competitive phage ELISA showed the inhibition with dose-dependent relationship. Conclusion ：The clone from phage-display peptide library could bind to p65 subunit of NF-κB specificaUy, and its sequence is recognized.

关 键 词：噬茵体肽库 NF-ΚB 酶联免疫吸附测定 

分 类 号：Q789[生物学—分子生物学]