Biotechnology Bulletin ›› 2013, Vol. 0 ›› Issue (9): 77-83.

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Cloning and Expression Analysis of Small GTP-binding Protein Gene from Dunaliella salina(DsRab)Under Salt Stress

Yu Zhujun Chai Xiaojie Zhang Xiaolin Zhang Ting Xue Fei   

  1. Key Laboratory of Mariculture & Stock Enhancement in North China’s Sea,Ministry of Agriculture,Key Laboratory of Marine Bio-resoursce Restoration and Habitat Reparation in Liaoning Province,Dalian Ocean University,Dalian 116023
  • Received:2013-04-15 Revised:2013-09-05 Online:2013-09-05 Published:2013-09-06

Abstract: Dunaliella salina small GTP-binding Protein Gene(GenBank Accession No.JN989548)was cloned by RT-PCR and RACE technology, named DsRab, the bioinformatics-analysis was performed, and the DsRab gene expression pattern was studied under 3 mol/L NaCl by Real-time quantitative RT-PCR method. The results showed that the full length of cDNA for DsRab Gene was 1 299 bp, which contains 78 bp 5'UTR, 609 bp 3'UTR and 612 bp ORF.The ORF codes a 203 amino acids protein with four GTP/GDP binding conserved domains, one effector domain, a cysteine residues at the COOH-terminus, and five common domain of Rab sub-family;Secondary structure prediction indicated that the α-helix, β-strands and random coil in it were 32.02% of, 23.65% and 44.33%. Protein homologue analysis showed that the DsRab shared high homology with Ypt/Rab from other species. The real-time quantitative PCR(RT-PCR)revealed that the expression of DsRab gene was increased by 4.9-fold under 3 mol/L NaCl comparising with that under normal level(including 1.5 mol/L NaCl)(P<0.01). Key words: Dunaliella salina Small GTP-binding protein gene RACE Bioinformatics-analysis QRT-PCR

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