Dugesia Casein Kinase 2¦Á Subunit And Mitogen-activated Protein Kinase Cdna Library Cloning And Bioinformatics Analysis

Dujesia japonica belonging to the phylum Nemertina, it is a model organism indevelopmentalbiology,regeneration biology and neurobiology. Because of its unique regenerative capacity and morphological plasticity,in recent years, molecular biology and functional genomics were used to study it. The mass of research on the planarian is about the mechanism of regeneration and molecular biology.The cDNA library of Dujesia japonica was constructed using SMARTTM cDNA Library Construction Kit,it was determined by PCR and RT-PCR.We separate and identify some genes from the cDNA library, bioinformatics tools were used to predict the gene function.we chose mitogen-activated proteinkinases and casein kinase 2 subunitαfrom the full length cDNA plasmid library.The full-length cDNA of mitogen-activated proteinkinases is 1315 bp, containing a 5'-untraslated region (5'-UTR) of 145bp. A 3'-UTR of 102 bp and a ployadenylation signal (AAAAAA). The content of GC is 38.48%.The open reading frame (ORF) of mitogen-activated proteinkinases is 1068bp, which encodes a deduced 355 amino acid peptide with a predicted molecular mass of 41359.5 and with the theoretical pI of 6.24.Sequence analysis showed that, mitogen-activated proteinkinases is hydrophilic Proteins, and mainly locates in the cytosol. It is stable.lt has no Signal Peptide and Transmembrane domain. Mitogen-activated proteinkinases belongs to PKc_like Super-family and it has 33 action sites,21 ATP binding sites,12 substrate binding sites and 21 KIM docking sites.The full-length cDNA of casein kinase 2 subunit a is 2004 bp. The open reading frame(ORF) of casein kinase 2 subunit is 1083 bp,which encodes a deduced 360 amino acid peptide with a predicted molecular mass of 41764.2 and with the theoretical pI of 6.43. The content of GC is 39.27%.Sequence analysis showed that, casein kinase 2 subunitαis hydrophilic Proteins, and mainly locates in the cytosol. It is unstable.It has no Signal Peptide and Transmembrane domain. Casein kinase 2 subunit abelongs to PKc_like Super-family and it has 26 action sites,20 ATP binding sites,11 substrate binding sites.The results of sequence alignments analysis by ClustalX showed that mitogen-activated proteinkinases shares 62%～66% identity with the other known mitogen-activated proteinkinases in animals. Casein kinase 2 subunit a shares 69%～74% identity with the other known Casein kinase 2 subunit ain animals. The similarity of two proteins'amino acid sequences in different species suggests similar molecular functions. The Phylogenetic tree construeted by NJ method showed that, mitogen-activated proteinkinases and casein kinase 2 subunit a clustered first with other Platyhelminthes proteins on the tree, and then with other mitogen-activated proteinkinases and casein kinase 2 subunit a from vertebrates, arthropoda,insects,and nematode.