缺失α1螺旋的Cry1Ac在大肠杆菌中的表达、纯化和活性分析Expression,Purification and Insecticidal Activity Analysis of Cry1Ac without Helix α-1
刘京国;郭蓓;赵晓萌;师光禄;
摘要(Abstract):
[目的]对缺失α1螺旋的Cry1Ac在大肠杆菌中的表达、纯化和活性进行分析。[方法]以苏云金芽孢杆菌HD-1中的质粒为模板,扩增缺乏α螺旋的Cry1Ac基因,并将其与pET28a载体连接,构建重组质粒。重组质粒转化并经IPTG诱导后,用SDS-PAGE检测目的蛋白的表达。提取目标蛋白,并涂在棉铃虫饲料表面,检测其杀虫活性。[结果]Cry1Acdelα1基因在大肠杆菌中能正常表达目标蛋白,但该目标蛋白是以包涵体形式存在。生侧结果显示,在相同浓度下,活化的Cry1Ac对棉铃虫的致死率达75%以上,而Cry1Acdelα1只有10%左右。[结论]用大肠杆菌表达的Cry1Acdelα1对棉铃虫基本没有杀虫活性。
关键词(KeyWords): Cry1Ac;α1片段;杀虫活性
基金项目(Foundation): 北京市自然科学基金(5092007和5112009)
作者(Author): 刘京国;郭蓓;赵晓萌;师光禄;
Email:
DOI: 10.13989/j.cnki.0517-6611.2012.06.004
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