@Article{taiwania1999444479, AUTHOR = {Yin-Chen Tzeng and Shu-Mei Pan}, TITLE = {Expression and Characterization of Rice Manganese Superoxide Dismutase in Escherichia coli}, JOURNAL = {Taiwania}, VOLUME = {44}, YEAR = {1999}, ISSUE = {4}, PAGES = {479-490}, URL = {https://taiwania.ntu.edu.tw/abstract/150}, ABSTRACT = {The corrected cDNA coding for rice mature MnSOD protein was made by PCR and inserted into pGEX-4T-1 expression vector. The recombinant DNA was transformed to E. coli BL21 and expression of GST-MnSOD fusion protein was induced by addition of IPTG to bacterial cultures. The homogeneous GST-MnSOD was purified by the one-step GST-glutathione affinity system. Both purified GST-MnSOD and recombinant MnSOD (rMnSOD) remained MnSOD activity, which showed an insensitivity towards KCN and H2O2. The molecular size of monomer of GST-MnSOD or rMnSOD was 50 kDa and 23 kDa respectively, and the functional form of both was dimer. The isoelectric point of rMnSOD was 4.64, but that of GST-MnSOD was in the range of pH 4.74 - 4.97. The SOD activities of GST-MnSOD and rMnSOD declined to 30 % after incubation at 60 ℃ for 20 minutes; but they were more stable in an alkaline pH environment.}, DOI = {10.6165/tai.1999.44(4).479} }