【摘要】 目的　构建耐辐射奇球菌 (D.radiodurans)锰超氧化物歧化酶 (Mn- SOD)基因的原核表达重组子 ,并在 E.coli BL 2 1(DE3)中表达。方法　用 PCR方法自 D. radiodurans基因组中扩增 Mn- SOD目的片段。将该基因与原核表达质粒载体 p ET- 30 a(+)连接 ,构建重组质粒 p ET- SOD,并转化表达宿主菌 E.coli BL 2 1(DE3)。用异丙基硫代 -β- D-半乳糖苷 (IPTG)诱导重组 SOD蛋白表达 ,SDS- PAGE分析表达产物。结果　获得了 D . radioduransMn- SOD基因的 p ET原核表达重组质粒 ,该质粒经 IPTG诱导能在 E.coli BL 2 1(DE3)中高效表达目的蛋白 ,蛋白活性可达 5 180 0 U / g湿菌体。结论　成功构建了原核表达重组质粒 p ET- SOD,实现了 SOD在原核细胞中的高效表达 ,表达产物活性较高 ,为重组 D. radiodurans Mn- SOD的进一步研究和应用奠定了基础。更多还原

【Abstract】 Objective To construct expressing recombinant of Mn-SOD of Deinococcus radiodurans and express the target protein in E.coli BL21(DE3). Methods SOD gene was amplified by PCR from genomic DNA of Deinococcus radiodurans and inserted into expression plasmid pET-30a(+) to create the recombinant pET-SOD. After being analyzed by the restriction endonuclease, the plasmid was transformed into E.coli BL21(DE3), and the recombinant protein was expressed after induction by the isopropyl-β-D-thiogalactopyranoside (IPTG) and was analyzed with SDS-PAGE. Results The recombinant plasmid pET-SOD was obtained, and the recombinant protein was highly expressed in E.coli BL21(DE3). The activity of recombinant superoxide dismutase was 51 800 U per gram of wet bacteria. Conclusion This study has provided a foundation for further studies and applications of the recombinant Mn-SOD.更多还原