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Int. J. Mol. Sci. 2010, 11(11), 4618-4630; doi:10.3390/ijms11114618

Molecular Cloning and Expression Analysis of P-Selectin from Zebrafish (Danio rerio)

1,2, 1, 2,* , 2 and 2
1 College of Life Sciences, Shandong Normal University, Jinan, Shandong 250014, China 2 Biology Institute of Shandong Academy of Sciences, Jinan, Shandong 250014, China
* Author to whom correspondence should be addressed.
Received: 21 October 2010 / Revised: 7 November 2010 / Accepted: 10 November 2010 / Published: 17 November 2010
(This article belongs to the Section Biochemistry, Molecular Biology and Biophysics)
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The glycoprotein P-selectin belongs to the selectin family of cell adhesion molecules. In this study, we cloned the full-length cDNA of P-selectin from zebrafish (Danio rerio) by the method of rapid amplification of cDNA ends polymerase chain reaction (RACE-PCR). Zebrafish P-selectin cDNA is 2,800 bp and encodes a putative 868 amino acid protein with a theoretical molecular weight of 122.36 kDa and isoelectric point of 6.27. A signal peptide of 25 amino acids is predicted at the N-terminus of the putative protein. All structural domains involved in P-selectin function are conserved in the putative protein. The amino acid sequence of zebrafish P-selectin is 37% to 39% identical to that of mammalian P-selectins. Real-time quantitative PCR and whole-mount in situ hybridization analysis revealed that P-selectin was expressed in early embryonic development, the expression increased from 0.2 hpf (1-cell stage) to 72 hpf, and the expression significantly upregulated within 30 minutes of ADP induction. The results indicate that the structure of P-selectin protein is highly conserved among species and zebrafish P-selectin plays an important role in early embryonic development and probably has similar biological function to mammalian P-selectins.
Keywords: P-selectin; zebrafish; cloning; expression; adenosine diphosphate P-selectin; zebrafish; cloning; expression; adenosine diphosphate
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Sun, G.; Pan, J.; Liu, K.; Wang, X.; Wang, S. Molecular Cloning and Expression Analysis of P-Selectin from Zebrafish (Danio rerio). Int. J. Mol. Sci. 2010, 11, 4618-4630.

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