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Oriental Galega is an excellent forage legumes , has a very important use value . Superoxide dismutase (SOD) is a widely played an important role the enzymes of eukaryotes metal , in stress resistance in plants . In this study, in order to explore the SOD gene Oriental Galega against oxidative damage , improve stress resistance impact RACE ( rapid amplification of cDNA ends ) method , according to the Oriental the Galega salt stress cDNA library of EST sequences , cloning of Cu / ZnSOD gene and its preliminary analysis . The Oriental Galega Cu / ZnSOD gene full-length cDNA sequences of 935 bp, 600 bp, encoding 199 amino acids of the open reading frame nucleotide sequence , the encoded protein molecular weight of 20.35 KDa the protein theoretical isoelectric point of 5.88 . Named the gene GoCu / ZnSOD the NCBI accession number HM777020 . Protein blast the procedures and ScanProsite server to analyze the amino acid sequence of the GoCu / ZnSOD Cu ion binding site is histidine 91,93,108 and 165, Zn ion binding site is histidine 108,116,125 and aspartic acid 128, GNAGGRIACGVV (183 -194 bp) , the C ( cysteine) may form a disulfide bond. Using real-time fluorescence quantitative PCR analysis GoCu / ZnSOD gene expression , tissue specificity results prove its expression was significantly higher than in the roots and stems of the leaves ; NaCl and PEG induced GoCu / ZnSOD gene expression levels are raised , low temperature for GoCu / ZnSOD gene expression , no significant effect of exogenous hormones ABA inhibition GoCu / ZnSOD gene expression . Build the subcellular localization of plant super expression vector pCAMBIA1302-GoCu/ZnSOD, and the use of the GFP protein under the excitation light can emit green fluorescence characteristics , tobacco injection method GoCu / ZnSOD protein analysis , laser scanning confocal microscopy results show , GoCu / ZnSOD protein located in the chloroplast .
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