作　　者： ; ; ;

机构地区： 广东药学院

出　　处： 《中国生化药物杂志》 2012年第4期329-332,共4页

摘　　要： 目的探讨重组家蝇天蚕素-人溶菌酶(Mdc-hly)在大肠杆菌中的表达条件及纯化。方法利用SDS-PAGE电泳和BandScan凝胶电泳图像分析系统研究诱导温度、诱导时机、异丙基硫代-β-D半乳糖苷(IPTG)诱导浓度和诱导时间对融合蛋白表达的影响。融合蛋白经His琼脂糖柱亲和色谱纯化后,Western blot鉴定。结果在起始菌浓度为A600=0.6时加入浓度为1.0 mmol/L的IPTG,37℃诱导6 h,融合蛋白的表达量最高,占菌体总蛋白的39.1%。纯化后融合蛋白纯度可达95%,Western blot鉴定为目的蛋白。结论确定了重组融合蛋白的最佳表达条件,并纯化了目的蛋白。 Purpose To optimize the expression conditions of recombinant Musca domestica cecropin-human lysozyme（Mdc-hly） and to purify recombinant protein.Methods Recombinant E.coli strains BL21（DE3） with plasmid pET-32a/Mdc-hly were induced at various temperatures,and various cell density with IPTG at different concentrations for different hours.Expressed protein was purified by chromatographic column and identified by Western blot.Results The expression level of fusion protein reached a peak value,accounting for 39.1% of the bacterial total protein,after the recombinant E.coli was induced with 1.0 mmol/L IPTG at 37 ℃,A600=0.6 for 6 h.SDS-PAGE and Western blot analysis indicated that the recombinant protein could be purified to 95% homogeneity.Conclusion The conditions for expression of fusion protein were preliminarily optimized,and the expressed protein was purified.

关 键 词： 家蝇天蚕素 溶菌酶 重组融合蛋白 表达条件 纯化

领　　域： [生物学]