[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.
Wang
Duochun
D
The Key Laboratory of Medical Molecular Bacteriology of the Ministry of Health, Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China. wangduochun@yahoo.com.cn
Kan
Biao
B
Gao
Shouyi
S
Liu
Yanqing
Y
D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D014734:Vibrio cholerae / Q000235:genetics / Q000378:metabolism / Q000821:virology*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D019151:Peptide Library; D010455:Peptides; D014764:Viral Proteins