Identification of oligopeptides binding to Mycoplasma hyorhinis P37 using a phage display library.
- Author:
Hua YANG
1
;
Qin FENG
;
Huiyu XU
;
Chengchao SHOU
Author Information
1. Department of Biochemistry and Molecular Biology, Cancer Hospital & Institute, Peking University, Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Beijing 100142, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Bacterial Proteins;
metabolism;
Base Sequence;
Molecular Sequence Data;
Mycoplasma hyorhinis;
metabolism;
Oligopeptides;
metabolism;
Peptide Library;
Protein Binding;
Swine
- From:
Journal of Biomedical Engineering
2011;28(6):1165-1188
- CountryChina
- Language:Chinese
-
Abstract:
Phage display random heptapeptide library was screened with recombinant P37 in this study. The positive phage clones were identified by ELISA and were sequenced, and the amino acid sequences of the polypeptides displayed on phage were deduced. After GST-polypeptides fusion protein was constructed and expressed, its binding to P37 was determined by GST-pull down and Western blot. After 4 rounds of bio-panning, the enriched positive phage clones were identified by ELISA. Eighteen positive phage clones were sequenced and the peptide sequences were as follows. ACAPKPPWLC (12/18), RPLSIDPWSPHL (3/18), RPLSNDPWSPHL (1/18), QNMMSPIEGVRI (1/ 18) and WAPEKDYMQLMK (1/18). The results from GST-pull down and Western blot showed that peptide RPLSIDPWSPHL could interact with P37. The study will be helpful for identifying the protein reacting with P37.
