Screening and identification of proteins that interact with receptor for advanced glycation end products (RAGE) via T7 select phage display system / 解放军医学杂志

ABSTRACT

Objective To screen and identify proteins that interact with receptor for advanced glycation end products (RAGE) with T7 select phage display system. Method GST-tagged fusion protein of RAGE was coated on glutathione suporflow resin and binding screens were performed with human lung T7 phage library. Results After 6 rounds of biopanning, 64 independent plaques were scraped and processed with EDTA. The inserted fragments in these plaques were amplified by PCR and the PCR products were purified by a gel recovery method. The sequences of the insertions were identified and analyzed with BLAST program in GeneBank. Twenty-five clones of encoding proteins were found. Conclusions T7 select phage display system is a convenient, rapid and effective method for screening binding proteins. This study provides an approach to explore complicated functions and novel mechanisms of RAGE-related signal transduction pathways.