Abstract：
In Triatoma infestans it was observed pyrethroid resistance attributed in part to an elevated oxidative metabolism mediated by cytochromes P450. The nicotinamide adenine dinucleotide phosphate (NADPH) cytochrome P450 reductase (CPR) plays a crucial role in catalysing the electron transfer from NADPH to all cytochrome P450s. The daily variations in the expression of CPR gene and a P450 gene (CYP4EM7), both associated with insecticide resistance, suggested that their expressions would be under the endogenous clock control. To clarify the involvement of the clock in orchestration of the daily fluctuations in CPR and CYP4M7 genes expression, it was proposed to investigate the effect of silencing the clock gene period (per) by RNA interference (RNAi). The results obtained allowed to establish that the silencing of per gene was influenced by intake schemes used in the interference protocols. The silencing of per gene in T. infestans reduced its expression at all the time points analysed and abolished the characteristic rhythm in the transcriptional expression of per mRNA. The effect of the per gene silencing in the expression profiles at the transcriptional level of CPR and CYP4EM7 genes showed the loss of rhythmicity and demonstrated the biological clock involvement in the regulation of t heir expression.
摘要：
在Triatoma感染中，观察到拟除虫菊酯抗性部分归因于细胞色素P450介导的氧化代谢升高。烟酰胺腺嘌呤二核苷酸磷酸（NADPH）细胞色素P450还原酶（CPR）在催化从NADPH到所有细胞色素P450的电子转移中起着至关重要的作用。CPR基因和P450基因（CYP4EM7）表达的日变异，两者都与杀虫剂抗性有关，建议他们的表达将在内生时钟控制下。为了阐明时钟参与CPR和CYP4M7基因表达的每日波动的编排，建议研究通过RNA干扰（RNAi）沉默时钟基因周期（每个）的效果。获得的结果允许确定每个基因的沉默受到干扰方案中使用的摄入方案的影响。在所分析的所有时间点，致病毛虫中每个基因的沉默都降低了其表达，并消除了每个mRNA转录表达的特征性节律。每个基因沉默在CPR和CYP4EM7基因转录水平的表达谱中的作用显示了节律性的丧失，并证明了生物钟参与了其表达的调节。
