Cloning and analysis of phosphoenolpyruvate carboxylase (PEPC) gene of Dunaliella salina

To investigate the function of phosphoenolpyruvate carboxylase(PEPC) gene of Dunaliella salina,a pair of degenerate primers was designed according to the conserved motifs of the PEPC of Chlamydomonas reinhardtii,Arabidopsis thaliana,and Arachis hypogaea.A cDNA fragment was obtained from green alga Dunaliella salina through RT-PCR,and then the full length of the cDNA was isolated by 3’ and 5’RACE.The isolated cDNA sequence was 3523 bp in length with a 2949 bp coding region that encoded 982 amino acid residues with the predicted relative molecular mass of 110560.5 dolton.In addition,homology analysis showed that PEPC of D.salina was highly similar to that of C.reinhardtii(69%),Chlorella variabilis(55%),Ostreococcus tauri(50%),and O.lucimarinus CCE9901(49%),suggesting that the cDNA isolated from Dunaliella salina was PEPC-encoding.