title: Molecular cloning and expression analysis of P-selectin from Zebrafish (Danio rerio).
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.
authors.affiliation: College of Life Sciences, Shandong Normal University, Jinan, Shandong 250014, China.
authors.forename: Guijin
authors.initials: G
authors.lastname: Sun
authors.forename: Jie
authors.initials: J
authors.lastname: Pan
authors.forename: Kechun
authors.initials: K
authors.lastname: Liu
authors.forename: Xue
authors.initials: X
authors.lastname: Wang
authors.forename: Sifeng
authors.initials: S
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017124:Conserved Sequence; D018507:Gene Expression Regulation, Developmental; D008969:Molecular Sequence Data; D019007:P-Selectin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D019007:P-Selectin; D012333:RNA, Messenger; D029961:Zebrafish Proteins