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棉花学报  2010 

棉铃虫HaHR3基因的克隆及其植物RNA干扰载体的构建

DOI: 1002-7807(2010)02-0157-06, PP. 157-162

Keywords: 棉铃虫,蜕皮调节转录因子,结构分析,RNA干扰

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Abstract:

采用RT-PCR方法克隆了棉铃虫蜕皮调节转录因子HaHR3基因,该基因含有1671bp的完整开放阅读框。序列分析表明,HaHR3与多种昆虫蜕皮调节转录因子高度同源。利用生物信息学方法对HaHR3的结构特征进行分析发现,HaHR3具有蜕皮调节转录因子超家族的典型特征,包括两个锌指结构和一个DNA结合结构域,不存在信号肽序列和N糖基化位点。选择HaHR3基因的部分片段构建了RNAi中间载体pRNAi1017-HaHR3sa,再将HaHR3正反向序列亚克隆至植物表达载体pCAMBIA2300-35S-OCS,成功构建了由35s启动子调控的HaHR3基因的正反向RNA干扰载体pCAM-RNAi-HaHR3。这一载体的成功构建为下一步通过植物介导的RNA干扰技术防治棉铃虫打下了坚实的基础。

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