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 w...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.展开更多
文摘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.
