叉头转录因子1基因FOXO1的特征及其在葱蝇夏滞育前期蛹体内糖脂代谢中的作用（英文）  

Characterization of fork head transcription factor 1 gene FOXO1 and its role in sugar and lipid metabolism in the summer prediapause pupae of Delia antiqua(Diptera: Anthomyiidae)

作　　者：徐燕玲 彭竹清 司风玲[1] 郝友进[1] XU Yan-Ling;PENG Zhu-Qing;SI Feng-Ling;HAO You-Jin(College of Life Science, Chongqing Normal University, Chongqing 401331, China)

机构地区：[1]重庆师范大学生命科学学院,重庆401331

出　　处：《昆虫学报》2018年第12期1384-1392,共9页Acta Entomologica Sinica

基　　金：重庆市科委项目(cstc2016jcyjA0152)

摘　　要：【目的】克隆和鉴定葱蝇Delia antiqua叉头转录因子1基因,探究其在葱蝇夏滞育前期蛹体内糖代谢和脂代谢中的作用。【方法】本研究基于葱蝇转录组数据利用3′RACE法克隆叉头转录因子1基因的全长开放阅读框;利用生物信息学法分析了该基因编码蛋白的序列特征、保守结构域和二级结构,并采用最大似然法对其与其他13种昆虫来源的同源序列进行了聚类分析。通过RNA干扰技术沉默目的基因后,采用实时定量PCR法分析葱蝇夏滞育前期蛹体内目的基因下游脂肪酶brummer基因(DaBmm)和磷酸烯醇式丙酮酸羧化酶基因(DaDepck)的表达规律,并对甘油三脂(TAG)、海藻糖和葡萄糖含量及总脂肪酶的活性变化进行分析。【结果】克隆得到了葱蝇叉头转录因子1基因DaFOXO1 (GenBank登录号:MG813258),其编码蛋白含有619个氨基酸,具有典型的叉头DNA结合域,核定位信号,2个14-3-3结合区和1个富含谷氨酰胺区;DaFOXO1与铜绿蝇Lucilia cuprina FOXO1蛋白的氨基酸序列有87%的一致性,并与其聚为一支。干扰葱蝇夏滞育前期蛹DaFOXO1后,8-20 h间可显著抑制DaBmm基因的表达,12-24 h间可显著影响TAG含量和总脂肪酶的活性,8-20 h(16 h除外)间可显著降低DaPepck的表达,但对葡萄糖和海藻糖的含量没有显著影响。【结论】本研究结果表明海藻糖和葡萄糖的积累在化蛹前已完成;DaFOXO1对DaBmm和DaDepck基因的表达调控可能有利于葱蝇夏滞育前期蛹体内的脂肪积累。同时也表明靶向Bmm依赖性脂类水解过程及其下游因子可为打破昆虫滞育提供一种有效策略。【Aim】To clone and identify fork head transcription factor 1 gene in the onion fly,Delia antiqua,so as to explore its roles in sugar and lipid metabolism in the summer prediapause pupae of this insect.【Methods】The full-length open-reading frame of fork head transcription factor 1 gene was cloned based on the transcriptome data of D.antiqua by 3′RACE method.The sequence characteristics,conserved domains and secondary structure of its encoded protein were analyzed using bioinformatic approaches.A maximum-likelihood phylogenic tree was constructed for the target protein and homologues from other thirteen insect species.After the target gene was silenced in the summer prediapause pupae of D.antiqua by RNA interference,the expression levels of its downstream genes including brummer gene(DaBmm)and phosphoenol-pyruvate carboxykinase gene(DaPepck)were measured by qPCR,and the contents of triacylglycerol(TAG),treaholse and glucose,and the total lipase activity were analyzed.【Results】The full-length cDNA sequence of the fork head transcription factor 1 gene DaFOXO 1 was cloned from D.antiqua,and deposited in the GenBank under the accession number MG813258.It encodes a 619-amino-acid protein with a typical fork head DNA binding domain,nuclear localization signal,two 14-3-3 binding motifs and a glutamine-rich domain.DaFOXO1 shares 87%sequence identity with FOXO1 of Lucilia cuprina at the amino acid level and they are clustered together.DaFOXO 1 knockdown in the summer prediapause pupae of D.antiqua significantly suppressed the expression of DaBmm between 8 h and 20 h after treatment,significantly affected the TAG content and total lipase activity between 12 h and 24 h after treatment,and significantly decreased the expression level of DaPepck between 8 h and 20 h(except 16 h)after treatment.Neither glucose content nor trehalose content in the ds DaFOXO 1-treated summer prediapause pupae of D.antiqua significantly changed.【Conclusion】The results indicate that the accumulation of trehalose and glucose has comp

关 键 词：葱蝇 夏滞育 RNA干扰 脂类代谢 糖代谢 

分 类 号：Q966[生物学—昆虫学]