Cloning and Bioinformatics Analysis of a DsMAPKKK Gene from Dunaliella salina

doi:10.11924/j.issn.1000-6850.2013-2740

Abstract

Abstract: To illustrat the function of MAPKs cascade reaction in salt-tolerance mechanism of Dunaliella salina, it was necessary to clone a MAPKKK gene which was a member of MAPKs signal transduction pathway and study its function.A mitogen-activated protein kinase kinase kinase gene was firstly cloned from Dunaliella salina by RT-PCR and RACE technology and named as DsMAPKKK(GenBank Accession No.KF366904) and further bioinformatics-analysis was conducted.The result showed that, full length of cDNA for DsMAPKKK gene was 1460 bp with a 879-bp open reading frame coding 292 amino acids.This protein was unstable, hydrophilic and located in cytoplasmic matrix without signal peptide and transmembrane region.The protein sequence contained a protein kinases ATP-binding region spanning from 24rd to 45rd amino acid and a Serine/Threonine protein kinase region spanning from 137rd to 149rd amino acid.Several potential phosphorylation sites and an important catalytic active site Asp141 were found in this protein.The main components of protein secondary structure were α-helix and random coil.A 3D model of protein tertiary structure was built successfully.Phylogenic analysis showed this protein had the closest evolutionary relationship with the corresponding proteins of Volvox carteri f. Nagariensis and Chlamydomonas reinhardtii.

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