水稻二化螟诱导表达基因OsSABATH启动子的克隆与缺失体构建

Cloning of the Promoter Sequence of Rice OsSABATH Gene Specifically Expressed by Chilo suppressalis Infestation and Construction of Its 5' Deletions

Northern杂交结果表明,OsSABATH基因受二化螟(Chilo suppressalis)为害诱导表达,但却不受机械损伤诱导。利用该基因序列和水稻基因组数据库相应序列设计引物,扩增到该基因编码区5′上游2050bp的启动子序列。用PLACE软件分析顺式作用元件,发现该基因起始密码子上游1.4kb的区域内存在几个可能与该基因表达调控相关的顺式作用元件,如W盒、DOF结合序列、GT-1、BIHD1结合序列等。将该基因启动子区域及其5′部分缺失构件与GUS报告基因相连,并克隆进pCAMBIA1391载体中,用于分析启动子活性及虫伤诱导特异性相关的顺式作用元件。

Northern blot analysis showed that the expression levels of the rice gene OsSABATH were up-regulated by infestation of rice striped stem borer, Chilo suppressalis, but not by mechanical wounding. A 2050-bp promoter sequence upstream of this gene was amplified using a pair of primers designed on the basis of the sequence of the OsSABATH and rice genome sequence database. With the aid of PLACE program, several cis-acting elements in the promoter sequence, including Wbox, DOF binding motif, GT-1 element, and BIHD1 binding motif, were recognized. For analyzing the activity of the promoter sequence and the functions of the herbivore feeding responsive cis-acting elements, the promoter region sequence and its 5 deletion constructs were fused to the GUS reporter gene in pCAMBIA1391 vector.