重组结核分枝杆菌异柠檬酸裂解酶表达特性的研究

Study on characteristics of recombinant Isocitrate Lyase expression from mycobacterium tuberculosis in E. coli

目的通过对所构建的重组结核分枝杆菌异柠檬酸裂解酶表达特性的研究,获取最佳可溶性表达。方法以IPTG诱导重组质粒pET28a(+)-icl和pET28a(+)-aceA在大肠杆菌BL21(DE3)和BL21(DE3)plysS细胞表达,筛选最佳诱导浓度、时间和温度。结果以0.25mmol/L和0.1mmol/L的IPTG在25℃诱导8h和6h后可溶性重组ICL蛋白和AceA蛋白的最高含量。结论本研究构建的结核分枝杆菌异柠檬酸裂解酶重组表达质粒,可获得可溶性表达蛋白,为抗结核药物的筛选奠定了基础。

Objective To obtain the soluble Isocitrate Lyase from Mycobacterium Tuberculosis H37Rv by optimizing the expression of recombinant ICL and AceA. Methods The expression of recombinant pET28a（ ＋ ） -icl and pET28a （ ＋ ） -aceA in E. coli BL -21 （DE3） and BL -21 （DE3）plysS were induced with IPTG in different concentration, time and temperature. Results The best induction of recombinant proteins of ICL was in 25℃ with 0.25mmol/L IPTG for 8h while in 25℃ with 0. 1mmol/L IPTG for 6h on aceA. Conclusion Obtaining the soluble expression of Isocitrate Lyase （ICL and aceA） from Mycobacterium Tuberculosis H37Rv can facihtate the discovery of antimicrobial agents against Mycobacterium Tuberculosis.