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

International Journal of Molecular Sciences

Guijin Sun¹  Jie Pan¹  Kechun Liu²  Xue Wang² 

[1] College of Life Sciences, Shandong Normal University, Jinan, Shandong 250014, China;Biology Institute of Shandong Academy of Sciences, Jinan, Shandong 250014, China

关键词: P-selectin;    zebrafish;    cloning;    expression;    adenosine diphosphate;   

DOI  :  10.3390/ijms11114618

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY   
© 2010 by the authors; licensee Molecular Diversity Preservation International, Basel, Switzerland.