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

Jianru Pan; Lijuan Chen; Huocong HE; Ying SU; Xiangling Wang; Xian LI; Cuihuang Chen; Lunqiao WU; Shutao Liu

Return

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

Author: Jianru PAN ¹ ; Lijuan CHEN ¹ ; Huocong HE ² ; Ying SU ² ; Xiangling WANG ¹ ; Xian LI ¹ ; Cuihuang CHEN ¹ ; Lunqiao WU ¹ ; Shutao LIU ¹
Author Information

1. College of Biological Science and Engineering, Fuzhou University, Fuzhou 350108, Fujian, China.
2. Laboratory of Radiation Oncology and Radiobiology, Fujian Cancer Hospital & Fujian Medical University Cancer Hospital, Fujian Key Laboratory of Tumor Translational Cancer Medicine, Fuzhou 350014, Fujian, China.
Publication Type:Journal Article
Keywords: Mn-SOD; construction; expression and purification; fusion protein; stability; transduction efficiency
From: Chinese Journal of Biotechnology 2017;33(7):1168-1177
CountryChina
Language:Chinese
Abstract: 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.