Abstract:To understand the function of alkaline phosphatase (ALP) in the development and metamorphosis of Paralichthys Olivaceus, full-length cDNA of ALP was cloned by the technique of RACE. The cDNA and deduced protein sequence were analyzed by bioinformatics. The results show that the fulllength cDNA sequence consists of 1 811bp, including an ORF encoding for 476 amino acids and a terminator codon. The GC content of coding region is different in selected ALPs. The predicted molecular weight of the deduced protein is 52 293.1 and the pI is 7.67. Molecular phylogeny tree shows that the ALP of Paralichthys Olivaceus shares high homology with the ALP of Tetraodon nigroviridis and Danio rerio. Several ALP functional sites, including the metal ion binding sites, phosphoserine intermediate, N-linked glycosylation sites, are conservative in those selected sequences. On tertiary structure prediction and structure alignment with human PALP, the flounder ALP has high similarity with that of human PALP, while 43% similarity is shared by the primary structure of them. It might possess one disulfide bond, which is composed by the cysteines 141 and 203. Few of the amino acids residues around the Zn and Mg metal ions are different between the above mentioned two kinds of ALP, including two similar amino acids, i.e. S155, T175.
陈晓武,施志仪. 牙鲆碱性磷酸酶cDNA序列分析与蛋白质高级结构预测[J]. 中国生物化学与分子生物学报, 2007, 23(06): 442-449.
CHENXiao-Wu,SHIZhi-Yi. cDNA Sequence Analysis and Tertiary Structure Prediction of Alkaline Phosphatase from Paralichthys olivaceus. Chinese Journal of Biochemistry and Molecular Biol, 2007, 23(06): 442-449.