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Updated: Aug 30, 2026
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Published on: December 1, 2020
[Screening and identification of proteins interacting with p38 MAP kinase via T7 phage-display screening system]
Zhi-jie Li1, Jing-hua Liu, Xiao-wei Gong
1Department of Pathophysiology and Key Laboratory of Shock and Microcirculation of PLA, First Military Medical University, Guangzhou 510515, China.
Objective:
To screen and identify proteins that interact with p38 mitogen-activated protein (MAP) kinases by means of T7 phage-display screening system.
Method:
His-tagged fusion protein of p38 MAP kinase was used to coat a 96-well ELISA plate and Ni-NTA resin, which served as the media for screening human lung and liver T7 phage cDNA libraries.
Results:
After 4 rounds of biopanning, 86 independent plaques were selected and processed by EDTA. The inserted gene fragments from these plaques were amplified by PCR, the products purified by a gel recovery method. The sequences of the insertions were identified and analyzed with BLAST program in GenBank. Forty-six clones were found to encode proteins.
Conclusion:
T7 phage-display screening system is convenient, rapid and effective for screening the P38 MAP kinase-binding proteins.
