期刊
PROTEIN JOURNAL
卷 32, 期 4, 页码 259-265出版社
SPRINGER
DOI: 10.1007/s10930-013-9482-5
关键词
Gene cloning; Sonneratia alba; Iron superoxide dismutase; Protein expression
资金
- National Natural Science Foundation of China [31170213, 91231106]
- Department of Education of Guangdong Province [2012KJCX0017]
In this study, a novel iron superoxide dismutase (FeSOD) gene from Sonneratia alba was cloned and then expressed in Escherichia coli Rosetta-gami, designated as SaFeSOD. The DNA sequence of SaFeSOD contained a 786-bp open reading frame which encodes a 261 amino-acid protein of 30.0 kDa. The 651-bp fragment coding for putative mature SaFeSOD was amplified and inserted into pET15b for expression. This recombinant SaFeSOD was subsequently isolated by Ni-trap column protein purification system. The apparent molecular mass of the purified enzyme was 25 kDa on SDS-PAGE. In comparison with FeSODs from other plant species, all iron-binding sites (His 27, His 80, Asp 164 and His 168) of SaFeSOD were conserved. SaFeSOD was found to have good pH stability in the pH range of 3.5-9.5 at 25 degrees C after 1 h incubation and was relatively stable and showed 78 % activity when incubated in 50 degrees C for 1 h. Quantitative real-time PCR experiments demonstrated that SaFeSOD was expressed in leaf, stem, flower, fruit and root tissues with the highest expression in leaf tissues.
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