Screening of binding proteins of HMGB1 promoter by phage display technique
- VernacularTitle:应用噬菌体展示技术筛选HMGB1启动子结合蛋白
- Author:
Ning DING
;
Hui XIAO
;
Ju GAO
;
Lixin XU
;
Shouzhang SHE
- Publication Type:Journal Article
- Keywords:
High mobility group box protein 1;
Promoter;
Phage display;
Peptide library;
Gene expression regulation
- From:
Chinese Journal of Pathophysiology
2010;26(1):28-31
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To screen the binding proteins to HMGB1 promoter by phage display technique. METHODS: HMGB1 promoter was incubated with phage display library. Unbound phages were eluted and phages bound to HMGB1 promoter were amplified. Twenty individual clones were randomly selected and identified by enzyme-linked immunosorbent assay (ELISA). Positive clones were characterized by DNA sequencing and the sequences were subjected for computer analysis. RESULTS: Positive phages binding to HMGB1 promoter were enriched after 4 rounds of biopanning. Twenty phage clones were selected and eleven clones of which were identified to bind specifically to HMGB1 promoter. The sequences in full length were obtained and searched for homologous sequences from GenBank. Altogether eight coding sequences were obtained, six of which were known proteins including activator protein-1(AP-1) and two of which were uncharacterized ones. CONCLUSION: Several proteins were obtained that bind specifically with HMGB1 promoter. The results will be useful for further studying the expression and regulation mechanism of HMGB1.
