应用噬菌体肽库筛选能封阻霍乱噬菌体VP1转染菌细胞的寡肽

噬菌体感染细菌首先要吸附于细菌表面受体 ,从目前报道的细菌与噬菌体相互作用的研究中发现 ,这些受体包括细菌细胞外膜上的蛋白、糖脂结构和鞭毛等。霍乱弧菌是霍乱的病原体 ,高守一等 (副霍乱资料汇编 ,1 984,2 37～ 2 4 5 .)从国内分离并选择出 5株噬菌体 (VP1～VP5 ) ,根据霍乱弧菌菌株对噬菌体的敏感性不同 ,将埃尔托型霍乱弧菌分为 32个噬菌体型。结合生物学分型方法 ,可区分埃尔托型霍乱弧菌的两类不同菌株 (流行株和非流行株 )和不同菌型。对各种来源的菌株进行分型 ,可作为一种追溯传染来源、传播途径和分析流行形式的流行病学研…

Screening for the Peptides Blocking Cholera Phage VP1 Transfection to Host Cell by Phage Random Amino Acid Peptide Library

Use Vibrio cholerae El Tor Typing Phage VP1 as ligand to screen phage-display random 7 amino acid peptide library, ELISA and inactivation experiment were used to identify positive clone. The ratio of output to input was increased after three rounds of screening. Pseudo-positive was decreased stepwise. It indicated the efficient enrichment. After three rounds of screening, 312 out of 360 phage clones were positive in ELISA, 1 clone shows blocking VP1 adsorbs to Vibrio cholerae by inactivation experiment. Sequencing result indicate that amino acid sequences are p245: LeuGlnGlnLysHisLeuLeu; p40,p55: GlnLeuIleMetIleArgHis; p69,p274 IleThrProArgAsnArgSer. Getting some peptides can mimick the VP1 receptor epitopes, one peptide can block VP1 transfection to host cell, it was a clue to study receptor's structure and phage infectious mechanism to host cell.