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Identification of Target Ligands of CORYNE in Arabidopsis by Phage Display Library
Author(s) -
Zhao Heng,
Li Shuzhen,
Sheng Jiping,
Shen Lin,
Yang Yuhui,
Yao Bin
Publication year - 2011
Publication title -
journal of integrative plant biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.734
H-Index - 83
eISSN - 1744-7909
pISSN - 1672-9072
DOI - 10.1111/j.1744-7909.2010.01028.x
Subject(s) - biopanning , meristem , biology , protein kinase domain , arabidopsis thaliana , phage display , kinase , arabidopsis , escherichia coli , biochemistry , microbiology and biotechnology , protein kinase a , peptide library , peptide , peptide sequence , gene , mutant
CORYNE (CRN) plays important roles in stem cell division and differentiation of shoot apical meristem (SAM) in Arabidopsis thaliana . The cytoplasmic kinase domain of CRN has been cloned and expressed in Escherichia coli , and further purified by two consecutive steps of affinity chromatography. By using this purified CRN as a ligand, a 12‐mer random‐peptide library was used to determine the specific amino acid sequences binding with the recombinant CRN molecule. After four rounds of biopanning, positive phage clones were isolated and sequenced, and further tested by enzyme linked immunosorbent assay for their binding ability and specificity. Two positive clones that specifically bind to the intracellular protein kinase domain of CRN have been identified. Alignment of these peptides and the kinase‐associated protein phosphatase (KAPP) shows high similarity, indicating that KAPP might interact with the cytoplasmic kinase domain of CRN and negatively regulate the CLV signal. Our current study would be helpful to better understand the CLV3 signal pathway.