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Cloning a glutathione peroxidase gene from Nelumbo nucifera and enhanced salt tolerance by overexpressing in rice.
- Source :
-
Molecular biology reports [Mol Biol Rep] 2014 Aug; Vol. 41 (8), pp. 4919-27. Date of Electronic Publication: 2014 Apr 09. - Publication Year :
- 2014
-
Abstract
- A full-length cDNA clone encoding an 866 bp-length glutathione peroxidase protein (NnGPX) was isolated from lotus (Nelumbo nucifera L.). The deduced amino acid sequence of the NnGPX gene had significant homology with ATGPX6. A 3D structural model of the NnGPX was constructed by homology modeling. The cloned NnGPX gene was expressed in Escherichia coli, and a fusion protein of about 40 kDa was detected after isopropyl thiogalactoside induction. Under different concentrations of Na2SeO3 treatments, NnGPX was found to be an enzyme that does not contain selenium. Real-time PCR analysis showed that the NnGPX gene was expressed in all organs of lotus, and its high expression mainly occurred in organs with active metabolisms. NnGPX transcript increased remarkably in response to cold, heat, mechanical damage, and salt treatment. Subsequently, the NnGPX gene was introduced in Oryza sativa cv. Yuetai B. PCR results verified the integration of this gene into the genome of rice and reverse transcription-PCR verified that this gene had been expressed in transgenic rice. The transgenic plants were significantly more tolerant to salt stress compared with the wild-type.
- Subjects :
- Amino Acid Sequence
Base Sequence
Cluster Analysis
Computational Biology
DNA Primers genetics
Gene Transfer Techniques
Molecular Sequence Data
Oryza physiology
Reverse Transcriptase Polymerase Chain Reaction
Sequence Analysis, DNA
Adaptation, Physiological genetics
Gene Expression Regulation, Plant genetics
Glutathione Peroxidase genetics
Models, Molecular
Nelumbo enzymology
Oryza genetics
Plants, Genetically Modified genetics
Salinity
Subjects
Details
- Language :
- English
- ISSN :
- 1573-4978
- Volume :
- 41
- Issue :
- 8
- Database :
- MEDLINE
- Journal :
- Molecular biology reports
- Publication Type :
- Academic Journal
- Accession number :
- 24715609
- Full Text :
- https://doi.org/10.1007/s11033-014-3358-4