柴化建;赵海泉;张丽君;罗焕亮;

• 1:深圳市职业技术学院
• 2:安徽农业大学生命科学学院
• 3:深圳出入境检验检疫局

摘要(Abstract)：

[目的]为提高植原体膜蛋白Imp基因在E.coli BL21(DE3)中的表达量,优化Imp基因的原核表达条件。[方法]通过设计正交试验,考察不同的培养条件对工程菌E.coli BL21(DE3)-pET-28a(+)-Imp的影响。在获得最佳培养条件的基础上考察不同诱导条件对Imp蛋白表达量的影响。利用SDS-PAGE和Gene Tools凝胶分析软件分析融合蛋白Imp的表达量。[结果]表达条件优化结果表明,最佳培养条件为:温度37℃,pH 7.0,装液量20%,振荡速度200 r/min;最佳诱导条件为:温度37℃,起始OD600≈1.5,IPTG终浓度0.1mmol/L,诱导培养时间6 h。[结论]在最佳条件下Imp表达量达70.98 mg/L,确定了Imp融合蛋白在大肠杆菌的优化表达条件。

关键词(KeyWords)： 植原体;Imp基因;表达条件;正交试验设计

Abstract:

Keywords:

基金项目(Foundation): 深圳市科技计划项目(JC200903180710A)

作者(Author): 柴化建;赵海泉;张丽君;罗焕亮;

Email:

DOI: 10.13989/j.cnki.0517-6611.2012.07.164

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