灰飞虱雌激素相关受体基因的克

发布时间：2018-05-05 22:55

本文选题：灰飞虱 + 雌激素相关受体　；参考：《昆虫学报》2017年03期

【摘要】：【目的】雌激素相关受体(estrogen-related receptor,ERR)是一类依赖配体激活的转录因子,能感受外源物质胁迫调控靶基因的转录,参与外源物质的代谢过程。本研究旨在通过克隆灰飞虱Laodelphax striatellus ERR基因,分析其序列和在氟啶虫胺腈胁迫下表达谱,探析其生理功能及在杀虫剂代谢中的作用。【方法】根据灰飞虱转录组数据信息,利用RT-PCR克隆灰飞虱ERR基因,并进行生物信息学分析;通过实时荧光定量PCR,分析暴露于氟啶虫胺腈(0.76 ng/头)后,灰飞虱4龄若虫ERR基因在不同时间点和不同组织中的表达量。【结果】从灰飞虱中克隆到ERR基因,命名为Ls ERR(Gen Bank登录号:KY210878),其c DNA序列全长1 854 bp,开放阅读框长1 260 bp,编码419个氨基酸,预测编码蛋白的分子量为47.70 k D。序列分析显示,Ls ERR具有核受体(nuclear receptors,NRs)家族成员的共同特征性结构:DNA结合区(DNA-binding domain,DBD)(第75-168位氨基酸)和配体结合区(ligand-binding domain,LBD)(第194-416位氨基酸)。采用APSSP2法预测Ls ERR蛋白二级结构,其中α螺旋占43.53%,β折叠占5.49%,无规则卷曲占50.98%。其中,在DBD区,有6个β折叠和3个α螺旋;在LBD区,有3个β折叠和11个α螺旋。系统发育分析表明,Ls ERR与褐飞虱Nilaparvata lugens ERR亲缘关系最近。灰飞虱4龄若虫暴露于氟啶虫胺腈后,12 h时其体内Ls ERR基因上调表达,24 h时达到表达高峰,48 h时表达量下调;Ls ERR在头部微弱表达,在腹部特异性高表达。【结论】Ls ERR是参与灰飞虱代谢氟啶虫胺腈的候选基因。本研究为解毒酶基因的调控和新分子靶标农药的研制提供分子基础。
[Abstract]:[objective] estrogen-related receptor ERR (ERR) is a ligand-activated transcription factor, which can regulate the transcription of target gene and participate in the metabolism of exogenous substance. The purpose of this study was to clone the Laodelphax striatellus ERR gene of planthopper, analyze its sequence and expression profile under flurazidonitrile stress, and explore its physiological function and its role in insecticide metabolism. [methods] according to the information of transcriptome data of fly planthopper, The ERR gene was cloned by RT-PCR and analyzed by bioinformatics. The expression of ERR gene of 4th instar nymph in different time points and in different tissues. [results] ERR gene was cloned from the plant planthopper. The c DNA sequence is 1 854 BP in length, and the open reading frame is 1 260 BP, encoding 419 amino acids. The predicted molecular weight of the encoded protein is 47.70 KD. Sequence analysis showed that the ERR had the common characteristic structure of the nuclear receptor NRs1 family members. The DNA binding domain of ERR was the 75-168 amino acids and the ligand binding domain of ligand binding domain LBDD (the 194-416 amino acid). The secondary structure of Ls ERR protein was predicted by APSSP2 method. 伪 helix accounted for 43.53%, 尾 fold for 5.49% and irregular crimp for 50.98%. Among them, there are 6 尾 folds and 3 伪 helix in DBD region and 3 尾 fold and 11 伪 helix in LBD region. Phylogenetic analysis showed that ERR had the closest relationship with Nilaparvata lugens ERR. The expression of Ls ERR gene in the 4th instar nymphs was down-regulated at 24 h after exposure to fludiconitrile and reached the peak at 48 h after exposure to fludiconitrile. [conclusion] Ls ERR is a candidate gene involved in the metabolism of fluazolidonitrile in planthopper. This study provides a molecular basis for the regulation of detoxifying enzyme gene and the development of new molecular target pesticides.
【作者单位】：江苏省农业科学院植物保护研究所;南京农业大学植物保护学院农作物生物灾害综合治理教育部重点实验室;淮北师范大学生命科学学院;
【基金】：江苏省自然科学基金项目(BK20150539) 国家科技支撑计划课题(2012BAD14B12-5) 江苏省农业科技自主创新资金(CX(15)1004) 农作物生物灾害综合治理教育部重点实验室开放课题基金(IMCDP201603)
【分类号】：Q78;S433

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