Expression, purification, stability and transduction efficiency of full-length SOD2 recombinant proteins].

Superoxide dismutase (SOD) family is necessary to protect cells from the toxicity of reactive oxygen species produced during normal metabolism. Among SODs, manganese-containing superoxide dismutase (Mn-SOD, SOD2) is the most important one. The DNA fragment containing the full nucleotide of full-length human SOD2 was synthesized and inserted into the prokaryotic expression vector pGEX-4T-1 with tag GST. DNA construct was then transformed into Escherichia coli BL21 (DE3) and expression was induced with IPTG at 25 ℃. The recombinant fusion protein GST-SOD2 (46 kDa) was purified from the bacterial lysate by GST resin column affinity chromatography. GST tag was cleaved with thrombin, and a crude SOD2 recombinant protein (25 kDa) was obtained and further purified by heparin affinity chromatography. Activities of the two SOD2 proteins were 1 788 and 2 000 U/mg, respectively. Both SOD2 proteins were stable under physiological condition and cell-penetrating (P<0.05). Our findings open the possibility to study the structure and effects of two full-length recombinant SOD2 proteins.

Citation

Jianru Pan, Lijuan Chen, Huocong He, Ying Su, Xiangling Wang, Xian Li, Cuihuang Chen, Lunqiao Wu, Shutao Liu. Expression, purification, stability and transduction efficiency of full-length SOD2 recombinant proteins]. Sheng wu gong cheng xue bao = Chinese journal of biotechnology. 2017 Jul 25;33(7):1168-1177

PMID: 28869736

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