HIV-1gp41核心结构模拟位的筛选与鉴定  

Screening and identification of HIV-1 gp41 core structure epitope from random phage display peptide library

作　　者：刘北一[1] 罗海波[1] 朱平[1] 姜世勃 富宁[1] 

机构地区：[1]第一军医大学基础部免疫教研室,广州510515 [2]纽约血液中心LFK研究所,纽约美国10021

出　　处：《中国免疫学杂志》2003年第7期461-463,472,共4页Chinese Journal of Immunology

基　　金：国家自然科学基金海外基金项目 ( 3 0 0 2 80 2 1)

摘　　要：目的 :筛选并鉴定HIV 1gp4 1核心表位。方法 :用识别HIV 1gp4 1的构象特异性单克隆抗体NC 1筛选噬菌体12肽库 ,通过夹心ELISA、NC 1特异性阻断实验、竞争抑制实验鉴定阳性噬菌体克隆 ,DNA序列分析阳性克隆。结果 :经 3轮筛选 ,随机挑取 2 4个噬菌体克隆 ,ELISA鉴定表明有 10个克隆可与NC 1结合 ,DNA序列分析并推导氨基酸序列 ,共 5种序列 :HDVHHRWVYLLS、ITVNEWLYTSEQ、HGRSHGMFKPKR、MGPIARPHWHLN、DMYRSPRPKPDT。其中gp4 1N肽和C肽所形成的复合物可特异性阻断表达HDVHHRWVYLLS ,VNEWLYTSEQ和MGPIARPHWHLN的克隆与NC 1的结合。结论 :所得序列HDVHHR WVYLLS ,VNEWLYTSEQ及MGPIARPHWHLN模拟HIV 1gp4 1六螺旋束核心表位。Objective:To identify and characterize the epitopes on core structure of HIV 1 gp41.Methods:A random phage displayed dodecapeptide library was screened with a conformation specific monoclonal antibody NC 1 specifically against the core structure of HIV 1 gp41.The positive clones were identified by sandwich ELISA,soluble NC 1 blocking assay and competitive inhibition assay.Results:After three rounds of screening,10 of 24 phage clones were identified as positive clones which can bind to NC 1.Amino acid sequences deduced from DNA sequences showed five different sequences:HDVHHRWVYLLS?ITVNEWLYTSEQ?HGRSHGMFKPKR?MGPIARPHWHLN?DMYRSPRPKPDT.The binding between phage clones(displayed HDVHHRWVYLLS,VNEWLYTSEQ and MGPIARPHWHLN, respectively)and NC 1 could be inhibited by N36 C34 complex.Soluble NC 1 could block the binding between phage clones and NC 1.Conclusion:The results indicate that HDVHHRWVYLLS,VNEWLYTSEQ and MGPIARPHWHLN are the mimotopes which could mimic the core structrue epitopes of six helix bundle of HIV 1 gp41.

关 键 词：HIV-1gp41 核心结构 模拟位 筛选 鉴定 

分 类 号：R392－33[医药卫生—免疫学]