Identification of a glucose-6-phosphate isomerase involved in adaptation to salt stress of <Emphasis Type="Italic">Dunaliella salina</Emphasis> |
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Authors: | Liuqing Cui Yurong Chai Jie Li Hongtao Liu Lei Zhang Lexun Xue |
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Institution: | (1) Institute of Tumor Molecular Surgery, The First Affiliated Hospital, Zhengzhou University, 40 Daxue Road, Zhengzhou, 450052, China;(2) College of Basic Medicine, Zhengzhou University, 100 Kexue Avenue, Zhengzhou, 450001, China;(3) Laboratory for Cell Biology, Department of Biology, Zhengzhou University, 100 Kexue Avenue, Zhengzhou, 450001, China;(4) Laboratory for Cell Biology, Zhengzhou University Medical College, 40 Daxue Road, Zhengzhou, 450052, China; |
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Abstract: | The unicellular green alga Dunaliella salina is a recognized model for studying plant adaptation to high salinity. To isolate some salt-induced proteins at proteomics
levels and to identify their expressions at gene levels, algal cells at logarithmic phase cultured in 1.5 and 3.5 M NaCl media
were harvested for protein extraction. Solubilized proteins were applied to two-dimensional gel electrophoresis (2-DE) and
analyzed by ImageMaster 2D Platinum software. Twenty-one protein spots whose intensities were elevated threefold to 13-fold
at 3.5 M NaCl as compared to 1.5 M NaCl were analyzed by matrix-assisted laser desorption/ionization tandem time of flight
mass spectrometry. One salt-induced protein isolated from the 2-DE gels was identified as a glucose-6-phosphate isomerase
(GPI) from D. salina (DsGPI). A full-length cDNA of DsGPI was obtained using rapid amplification of cDNA end technique, and it was shown by heterologous
expression to encode a protein with a molecular weight consistent with the protein spot in the 2-DE gels. Real-time quantitative
RT-PCR demonstrated that the mRNA of DsGPI was induced up to eightfold (P < 0.01) by 2.5 M and 14-fold higher (P < 0.01) by 3.5 M NaCl than by 1.5 M NaCl, respectively. It is concluded that the protein isolated through 2-DE is indeed
DsGPI and that the DsGPI gene may be involved in adaptation to high salinity. |
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