[Screening and identification of proteins interacting with p38 MAP kinase via T7 phage-display screening system].
OBJECTIVE
To screen and identify proteins that interact with p38 mitogen-activated protein (MAP) kinases by means of T7 phage-display screening system.


METHODS
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.


CONCLUSIONS
T7 phage-display screening system is convenient, rapid and effective for screening the P38 MAP kinase-binding proteins.
Li
Zhi-jie
ZJ
Department of Pathophysiology and Key Laboratory of Shock and Microcirculation of PLA, First Military Medical University, Guangzhou 510515, China.
Liu
Jing-hua
JH
Gong
Xiao-wei
XW
Qin
Qing-he
QH
Huang
Hao
H
Deng
Peng
P
Wang
Jing-zhen
JZ
Zhao
Shan-chao
SC
Sun
Xue-guang
XG
Liu
Ya-wei
YW
Zhao
Ke-shen
KS
Jiang
Yong
Y
D017123:Bacteriophage T7 / Q000235:genetics*; D015723:Gene Library; D006801:Humans; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism*; D011485:Protein Binding; D048051:p38 Mitogen-Activated Protein Kinases
D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases