HPV 58型E7蛋白特异结合多肽的筛选及分析  

Screening and analysis peptides that bind specifically to HPV 58 E7 protein

作　　者：范久松[1,2] 赵洪彬[3] 林洁[3] 马岚[2] 

机构地区：[1]清华大学生命科学学院,北京100084 [2]清华大学深圳研究生院生命与健康学部,广东深圳518055 [3]云南大学生命科学学院,云南昆明650091

出　　处：《云南大学学报（自然科学版）》2011年第5期615-620,共6页Journal of Yunnan University(Natural Sciences Edition)

基　　金：云南大学理(工)科校级科研项目资助(2009C13R)

摘　　要：利用M13噬菌体展示技术筛选HPV 58型E7蛋白特异性结合12肽,对这些多肽的序列比对分析得出其共有序列,同时通过BLASTP序列对比分析获得能与E7结合的内源蛋白.利用GST-E7融合表达蛋白为正筛靶分子,GST蛋白为负筛靶分子进行4轮的正负亲和筛选,经过ELISA活性鉴定获得71株阳性克隆,通过DNA测序和序列分析,得到2个能与E7蛋白发生特异性结合的共有序列NHXXANPQQXPQ和TMGFTAPRF-PHY.通过BLASTP分析所有71个多肽在体内的同源蛋白,它们能为对E7蛋白的致癌机理研究提供方向.By using M13 phage display techniques,12-mer peptide ligands that bind specifically to HPV 58 E7 protein were isolated,and then we compared all the sequences to get consensus sequences,moreover,through BLASTP endogenous proteins which probably interact with E7 protein were found.Using GET-E7 fusion protein as a positive screening target and GST protein as a negative one,we took four rounds of negative-positive selection,after that,71 positive clones were selected.By DNA sequencing and sequence analysis,we got two consensus sequences：NHXXANPQQXPQ and TMGFTAPRFPHY.We analyzed the homologous proteins of the 71 peptides by BLASTP,which will be very helpful in the study of E7 carcinogenesis.

关 键 词：噬菌体展示 E7蛋白 多肽 

分 类 号：Q786[生物学—分子生物学]