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.
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.
- Content Type Journal Article
- DOI 10.1007/s10811-009-9494-x
- Authors
- Liuqing Cui, Zhengzhou University Institute of Tumor Molecular Surgery, The First Affiliated Hospital 40 Daxue Road Zhengzhou 450052 China
- Yurong Chai, Zhengzhou University College of Basic Medicine 100 Kexue Avenue Zhengzhou 450001 China
- Jie Li, Zhengzhou University Laboratory for Cell Biology, Department of Biology 100 Kexue Avenue Zhengzhou 450001 China
- Hongtao Liu, Zhengzhou University Laboratory for Cell Biology, Department of Biology 100 Kexue Avenue Zhengzhou 450001 China
- Lei Zhang, Zhengzhou University Laboratory for Cell Biology, Department of Biology 100 Kexue Avenue Zhengzhou 450001 China
- Lexun Xue, Zhengzhou University Institute of Tumor Molecular Surgery, The First Affiliated Hospital 40 Daxue Road Zhengzhou 450052 China
- Journal Journal of Applied Phycology
- Online ISSN 1573-5176
- Print ISSN 0921-8971
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