J Integr Plant Biol. ›› 2011, Vol. 53 ›› Issue (4): 281-288.DOI: 10.1111/j.1744-7909.2010.01028.x

• Plant-environmental Interactions • Previous Articles     Next Articles

Identification of Target Ligands of CORYNE in Arabidopsis by Phage Display Library

Heng Zhao1*, Shuzhen Li2, Jiping Sheng2, Lin Shen2, Yuhui Yang1 and Bin Yao1*   

  1. 1Key Laboratory for Feed Biotechnology of the Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China
    2College of Food Sciences, China Agricultural University, Beijing 100080, China

Abstract:

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.

Zhao H, Li S, Sheng J, Shen L, Yang Y, Yao B (2011) Identification of target ligands of CORYNE in Arabidopsis by phage display library. J. Integr. Plant Biol. 53(4), 281–288.

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