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《Molecular Plant Breeding》 2018-10
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Cloning and Expression Vectors Establishment of Plasma Membrane Na~+/H~+ Antiporter Gene(RcSOS1) in Castor(Ricinus communis L.)

Li Ping;Wang Xiaoyu;Xu Hui;Wang Xuejuan;Zhang Jixing;College of Life Science, Inner Mongolia University for Nationalities;Inner Mongolia Industrial Engineering Research Center of Universities for Castor;Inner Mongolia Key Laboratory of Castor Breeding;Inner Mongolia Collaborative Innovation Cultivate Center for Castor;Horqin Plant Stress Biology Research Institute of Inner Mongolia University for Nationalities;  
Degenerate primers were designed according to the conservative region of SOS1(NCBI: KX943304.1)gene in other species. The RcSOS1 gene sequence was cloned by the method of RT-PCR and RACE. The open reading frame was 3 432 bp, and the deduced protein consisted of 1 143 amino acid residues, which had the highest homology with SOS1 gene in Jatropha carcas(80.4%). Also, it encoded a Na~+/H~+antiporter. Hydrophobicity analysis showed that the RcSOS1 protein was predicted to contain 12 transmembrane domains at the N-terminal part and a long hydrophilic cytoplasmic tail at the C-terminal. The analysis data of the second structure of RcSOS1 indicated that α-helix and random coil were the largest structural elements followed by extension chain and β-sheet. The tertiary structure analysis of RcSOS1 indicated that it was a Na~+/H~+antiporter on the plasma membrane. Full length amplification primer with Bam H Ⅰ and Sal Ⅰ loci was designed by RcSOS1 full length sequence to amplify the full length. The positive expression vector of RcSOS1 gene was successfully constructed by the combination of Bam HⅠ and Sal Ⅰ after double enzyme digestion and expression vector p CG-1301, which could provide help for the further study on the function of RcSOS1 gene and its role in the regulation of salt tolerance.
【Fund】: 国家自然基金项目(31760399;31260336;31701467)资助
【CateGory Index】: Q943.2;S565.6
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