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Functional Selection of Peptide That Specifically Inhibits Salmonella Typhi Adhesion to/invasion of Human Monocytic Cells by Ribosome Display

Author: WuHongYan
Tutor: ZhangXiaoLian
School: Wuhan University
Course: Immunology
Keywords: Ribosome display technology Salmonella typhi The IVB type cilia Binding polypeptides
CLC: R392
Type: Master's thesis
Year: 2005
Downloads: 142
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Abstract


More than two thousand kinds of Salmonella , Salmonella typhi is important pathogens, only human pathogenic and cause typhoid typhoid . Salmonella typhi can invade the intestinal cells to survive in macrophages and dendritic cells . Discovered in recent years Salmonella typhi pathogen virulence island IVB cilia and typhoid bacillus closely related to human - specific pathogenic but the IVB cilia newly discovered human monocytes role of the mechanism is unclear . For further study of typhoid bacillus the IVB type cilia function and the development of human mononuclear cells can specifically inhibit the invasion of Salmonella typhi antimicrobial polypeptide preparation , the present experiment in vitro synthesized primers and oligonucleotides containing 36 random sequence by two round of PCR to construct a library of random DNA , followed by in vitro coupled transcription and translation , ribosomal library containing random peptide 12 ribosomal library constructed , screened with a 12 peptide the typhoid bacillus IVB - the cilia structural proteins prePilS binding R ( RQERSSLSKPVV ) . The invasion inhibition experimental results show that The peptide R can inhibit the cilia Salmonella typhi invasion of human monocytes Department of THP - 1 has no effect on non-ciliated Salmonella typhi , the same size and solubility of 12 peptide R 12 peptide Y ( as a control polypeptide ) ciliated and non-ciliated Salmonella typhi invasion of cells had no inhibitory effect ; isothermal titration calorimetry experiments ( ITC ) found of 12 peptide R prePilS protein Ka value 0.4 × 10 5 L · mol -1 ~ 2.2 × 10 5 L · mol -1 , showed that 12 the peptide R prePilS protein with high affinity . In this study, for the first time in the international screening and to determine the specific binding of Salmonella typhi IVB cilia 12 peptide R , screened 12 peptides R has the potential for resistance to Salmonella typhi infection of human cells , the data in this article for further understanding of Salmonella typhi IVB cilia pathogenic mechanisms laid the foundation.

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