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Characterization of a cDNA coding for an extracellular calmodulin-binding protein from suspension-cultured cells of Angelica dahurica

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Abstract

In order to characterize a specific extracellular 21-kDa calmodulin-binding protein (named: ECBP21) from Angelica dahurica L. suspension-cultured cells, the cDNA coding for the protein has been cloned. Here, Southern blot analysis shows that there are at least two copies of ECBP21 gene in Angelica genome. Using truncated versions of ECBP21 and synthetic peptide in CaM binding assays, we mapped the calmodulin-binding domain to a 16-amino acid stretch (residues 200–215) at the C-terminal region. The ECBP21 was localized in the cell wall area by the immunogold electron microscopy and by GFP labeling method. These results define ECBP21 as a kind of an extracellular calmodulin-binding protein (CaMBP). Furthermore, using Northern blot analysis, we examined the expression dynamics of ecbp21 during the incubation of Angelica suspension-cultured cells and the treatments with some growth regulators. The above studies further provide the molecular evidence for the existence of the gene coding for extracellular CaMBPs and imply a possible role for ECBP21.

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Abbreviations

ABA:

Abscisic acid

CaM:

Calmodulin

CaMBD:

Calmodulin-binding domain

CaMBP:

Calmodulin-binding protein

ECBP21:

21-kDa extracellular CaMBP

GA3:

Gibberellin A3

GFP:

Green fluorescent protein

JA:

Jasmonic acid

RACE:

Rapid amplification of cDNA ends

SA:

Salicylic acid

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Acknowledgements

We thank Dr. Haseloff J. for his generous gift of the pBIN 35S-mGFP4 plasmid. This research was supported by the National Natural Science Foundation of China (Grant No. 30170476), the National Key Basic Research Special Funds of China (Grant No. G19990117) and the National Key Research Funds of China (Grant No. 90208004).

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Authors and Affiliations

  1. Institute of Molecular Cell Biology, College of Life Sciences, Hebei Normal University, Shijiazhuang, Hebei, 050016, People’s Republic of China

    Guo-Hong Mao, Li-Xia Hou, Cun-Bao Ding, Su-Juan Cui & Da-Ye Sun

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  1. Guo-Hong Mao
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  2. Li-Xia Hou
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  3. Cun-Bao Ding
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  4. Su-Juan Cui
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  5. Da-Ye Sun
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Correspondence to Da-Ye Sun.

Additional information

G.-H. Mao and L.-X. Hou contributed equally to this work.

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Mao, GH., Hou, LX., Ding, CB. et al. Characterization of a cDNA coding for an extracellular calmodulin-binding protein from suspension-cultured cells of Angelica dahurica. Planta 222, 428–437 (2005). https://doi.org/10.1007/s00425-005-1558-9

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  • DOI: https://doi.org/10.1007/s00425-005-1558-9

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