从HCV核心蛋白噬菌体随机展示肽库中筛选抗原表位  

Screening antigenic epitopes from HCV core protein random peptide libraries displayed on phage

作　　者：潘卫[1] 戚中田[1] 吴晓兰[1] 贺祥[1] 潘欣[1] 陈秋莉[1] 杜平[1] 

机构地区：[1]第二军医大学微生物学教研室,上海市翔殷路800号上海200433

出　　处：《细胞与分子免疫学杂志》2001年第1期20-23,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金重点资助!No.39830330;上海市科技发展基金资助!No.98JC14023

摘　　要：目的探讨病毒抗原序列研究的新方法。方法应用噬菌体展示技术，筛选ＨＣＶ核心蛋白的抗原序列。用抗－ＨＣＶ核心抗体单独阳性的血清，对巳构建的ＨＣＶ核心蛋白噬菌体随机展示肽库进行４轮筛选，检测筛选阳性的克隆数、插入阳性率和杂交阳性率来评价筛选的效果。测定和分析７个杂交阳性克隆的ＤＮＡ序列。结果所测定的７个序列中，６个为ＨＣＶ核心蛋白序列，其中５个被正确地展示于噬菌体表面，１个可能被正确展示，这些序列均含有重要的ＨＣＶ核心抗原表位。另１个为大肠杆菌ｎｒｆａ基因。结论噬菌体展示技术完全可应用于病毒蛋白抗原序列的筛选，并具有简便、快速、准确的优点。Aim To screen out the antigenic sequences from HCV core protein random peptide libraries displayed on phage and to explore a new way to screen the viral antigens. Methods The anti-HCV core antibody-positive serum was used to screen antigenic peptides from the HCV core protein random peptide libraries displayed on phage for 4 rounds. Detection of numbers of positive clones, positive rate of insertion of HCV random DNA and positive rate of hybridization with HCV core probes were used to evaluate the screening effects. The DNA sequences of 7 selected clones with positive hybridization were determined and analysed. Results Six out of 7 sequences are HCV core protein sequences, in which 5 were perfectly displayed, and one was possibly displayed. These sequences included several major HCV core antigenic epitopes. The remaining one was E. coil nr-fa gene. Conlusion The phage display technique can be applied to study the viral antigenic peptides with the advantages of simple, accuracy and rapidity.

关 键 词：丙型肝炎病毒 核心蛋白 噬菌体 随机肽库 抗原 

分 类 号：Q78[生物学—分子生物学] R392.11[医药卫生—免疫学]