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Screening and identification of proteins interacting with p38 MAP kinase via T7 phage-display screening system



Screening and identification of proteins interacting with p38 MAP kinase via T7 phage-display screening system



Di 1 Jun Yi Da Xue Xue Bao 23(11): 1131-1133



To screen and identify proteins that interact with p38 mitogen-activated protein (MAP) kinases by means of T7 phage-display screening system. 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. 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. T7 phage-display screening system is convenient, rapid and effective for screening the P38 MAP kinase-binding proteins.

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Accession: 050262706

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PMID: 14625169


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