棉铃虫细胞色素P450基因CYP9A17v2核心启动子区缺失分析  

Deletion analysis of the core promoter region of CYP9A17v2 from Helicoverpa armigera(Lepidoptera:Noctuidae)

作　　者：严宇澄[1] 武淑文[1] 杨亦桦[1] 吴益东[1] 

机构地区：[1]南京农业大学植物保护学院,农业部病虫害监测与防控重点开放实验室,南京210095

出　　处：《昆虫学报》2009年第8期825-831,共7页Acta Entomologica Sinica

基　　金：国家自然科学基金项目(30471154)

摘　　要：CYP9A17v2的过量表达与棉铃虫Helicoverpa armigera对拟除虫菊酯类杀虫剂的抗性相关。为了研究CYP9A17v2表达调控机制,对CYP9A17v2核心启动子区域的功能进行了分析;构建了含荧光素酶报告基因和CYP9A17v2启动子不同长度缺失片段(-1095～+43)的重组质粒,转染Sf9细胞瞬时表达后用双荧光素酶报告基因检测系统检测启动子活性。功能分析结果表明:所有的7个缺失片段均具有启动子活性,-197～+43启动子区域的转录活性最高。在CYP9A17v2基因5′-调控区-197～-113区域内可能存在转录增强因子的结合位点,而在-1095～-197区域内可能存在转录抑制因子的结合位点。本研究为探索棉铃虫CYP9A17v2过量表达的转录调控机理奠定了重要基础。Overexpression of cytochrome P450 CYP9A17v2 gene is involved in pyrethroid resistance in Helicoverpa armigera （Htibner）. In order to study the mechanisms underlying regulation of CYP9A17v2, functional analysis of the promoter of CYP9A17v2 from H. armigera was performed in this study. Luciferase reporter plasmids containing serially truncated promoter fragments （ - 1 095 - ＋ 43 ） of CYP9A17v2 were transiently transfected into Sf9 cells, and the promoter activity was measured with dual-luciferase reporter assay system. Functional analysis showed that all the seven deleted fragments had promoter activity, and a region between - 197 and ＋ 43 exhibited the highest level of promoter activity. Transcription enhancer elements may exist in the region between - 197 and - 113 of CYP9A17v2 5＇-regulation area, and transcription repressor elements may situate in the region between - 1 095 and - 197. This study would provide an important foundation for investigating transcriptional regulation mechanisms of CYP9A17v2 overexpression in H. armigera.

关 键 词：棉铃虫 细胞色素P450 CYP9A17v2 启动子 双荧光素酶报告基因系统 转录调控 

分 类 号：Q965.9[生物学—昆虫学]