摘要
为对禽流感病毒(AIV)M1蛋白表(拟)位进行分析,本研究采用针对AIV M1蛋白的型特异性单克隆抗体(MAb),淘选M13噬菌体展示的7肽随机肽库,进行M1蛋白表(拟)位分析。筛选获得共有序列MDRxL或HPR,定位于M1蛋白93～99位(93MDRAVKL99)和222～230位(222HPNSSAGLR230)氨基酸区域。采用ELISA、竞争性ELISA分析不同噬菌体拟位与抗M1的MAb免疫反应性,表明含有MDRxL或HPR基序的噬菌体拟位能够与MAb发生特异性结合,并且其结合能够被天然病毒抗原抑制或阻断,表明拟位多肽真实模拟病毒蛋白上与MAb结合的抗原决定簇或表位,提示M1蛋白93～99位(93MDRAVKL99)和222～230位(222HPNSSAGLR230)氨基酸区域构成AIV型特异性表位。
Matrix protein 1 （M1） of avian influenza virus （AIV） is composed of 252 amino acid residues which is one of the basis for typping of influenza virus. To identify the epitope in M1 of AIV, a 7-mer random peptide phage display library was screened for epitope mapping with the monoclonal antibody （MAb） against A1V M1 protein. The consensus sequence of MDRxL or HPR was recognized by the MAb within 93MDRAVKL99 and 222HPNSSAGLR230 of M1 protein. Furthermore, Using ELISA and competitive ELISA to analysis immunoreactivities of different phage mimotopes to the MAb, the results showed the mimotopes with basic amino acid sequence of MDRxL or HPR was able to specifically reacted with the MAb which was blocked by natural virus antigen. It demonstrated that the mimotope peptides truly mimicked the MAb binding determinants or linear epitope on the viral protein. This result indicated that the sequences of 93MDRAVKL99 and 222HPNSSAGLR230 in M1 protein constituted the influenza type-specific epitope.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2012年第9期732-735,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
云南省应用基础研究项目(2008ZC071M)
云南省后备人才基金项目(2009CI061)
云南省高层次科技人才培引工程(20080C002)
关键词
禽流感
单克隆抗体
M1蛋白
表位
定位
avian influenza
monoclonal antibody
matrix protein 1
epitope
mapping
