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Molecular Cloning and Expression Analysis of P-Selectin from Zebrafish (Danio rerio)
College of Life Sciences, Shandong Normal University, Jinan, Shandong 250014, China
Biology Institute of Shandong Academy of Sciences, Jinan, Shandong 250014, China
* Author to whom correspondence should be addressed.
Received: 21 October 2010; in revised form: 7 November 2010 / Accepted: 10 November 2010 / Published: 17 November 2010
Abstract: 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
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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.
Sun G, Pan J, Liu K, Wang X, Wang S. Molecular Cloning and Expression Analysis of P-Selectin from Zebrafish (Danio rerio). International Journal of Molecular Sciences. 2010; 11(11):4618-4630.
Sun, Guijin; Pan, Jie; Liu, Kechun; Wang, Xue; Wang, Sifeng. 2010. "Molecular Cloning and Expression Analysis of P-Selectin from Zebrafish (Danio rerio)." Int. J. Mol. Sci. 11, no. 11: 4618-4630.