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- Title
海参铜锌超氧化物歧化酶原核表达载体的构建、表达及活性鉴定.
- Authors
车明月; 穆贤; 李学一; 张齐; 丛丽娜; 牛庆昌; 李成
- Abstract
Apostichopus japonicus Cu/Zn-superoxide dismutase(Aj-SOD1) with activity was successfully expressed in prokaryotic host E.coli BL21(DE3).Total RNA was extracted from the intestinal tissue of Apostichopus japonicus by Trizol.Cu/Zn-SOD1 gene was amplified by RT-PCR to obtain the coding gene sequence of Aj-SOD1(GenBank:JX097096.1,459 bp)and pMD18-Aj-SOD1 was constructed.Cloning vector of pMD18-Aj-SOD1 with restriction sites of Nde Ⅰ/Xho Ⅰ was constructed. The recombinant expression vector of pET30 a-Aj-SOD1 was constructed with pET30 a(+)double enzyme digested by Nde Ⅰ/Xho Ⅰ and Aj-SOD1.pET30 a-Aj-SOD1 was transformed into E.coli BL21(DE3)competent cells and Kan+resistance screening positive transformants were selected.Aj-SOD1 proteins with antioxidant activity were successfully obtained by inducing with0.1 mmol/L IPTG after 8 hand detected by SDS-PAGE and pyrogallol autoxidation method.
- Publication
Journal of Dalian Polytechnic University, 2018, Vol 37, Issue 1, p5
- ISSN
1674-1404
- Publication type
Academic Journal
- DOI
10.19670/j.cnki.dlgydxxb.2018.0102