柞蚕蛹解除滞育过程中海藻糖合成酶基因的表达变化

发布时间：2018-02-27 16:29

本文关键词： 柞蚕滞育海藻糖合成酶基因表达滞育解除　出处：《昆虫学报》2016年09期 　论文类型：期刊论文

【摘要】：【目的】克隆柞蚕Antheraea pernyi海藻糖合成酶(trehalose-6-phosphate synthase,TPS)基因,并对其进行组织表达分析,探讨该基因在柞蚕滞育蛹解除滞育过程中的表达规律,为阐明柞蚕滞育期间碳水化合物代谢规律与蛹滞育解除的关系提供数据支持。【方法】利用PCR及3'RACE技术从柞蚕幼虫脂肪体组织中克隆得到TPS基因,并进行生物信息学分析;RT-PCR检测该基因在柞蚕幼虫各组织中的表达分布,进一步采用Real-time PCR分析柞蚕滞育蛹解除滞育过程中该基因在脂肪体组织和血淋巴中的表达量变化。【结果】克隆获得柞蚕海藻糖合成酶基因并命名为ApTPS。其开放阅读框长2 487 bp,编码828个氨基酸,蛋白预测分子量为93.19 k D,等电点(p I)4.61;无信号肽,无跨膜区。蛋白质亚细胞定位预测该蛋白定位于细胞质中;蛋白质结构域分析表明,ApTPS有两个保守功能区:TPS(第22-497位氨基酸)和TPP(第532-772位氨基酸)。组织特异性分析表明,ApTPS基因在柞蚕幼虫脂肪体中表达量最高;柞蚕解除滞育过程中,ApTPS在脂肪体和血淋巴中的表达量均有所升高,且显著高于对照组(P0.05),但血淋巴中表达量的升高滞后于脂肪体。【结论】结果提示ApTPS参与了柞蚕蛹滞育中碳水化合物代谢调控并在其中发挥重要作用,与柞蚕蛹滞育解除关系密切。
[Abstract]:[Objective] cloning of trehalose synthase (trehalose-6-phosphate pernyi Antheraea from synthase, TPS) gene and its tissue expression analysis, explore the gene expression of diapause termination in Antheraea pernyi diapause pupae, to provide data support for the relationship during the clarify of Antheraea pernyi diapause carbohydrates metabolism and pupal diapause termination. [method] cloned from Tussah larvae in the fat body tissue TPS gene was obtained by using PCR and 3'RACE technology, and bioinformatics analysis; distributed RT-PCR detection expression of the gene in different tissues in Tussah larvae, further using Real-time PCR analysis of Antheraea pernyi diapause pupae diapause of the gene in fat body and hemolymph of expression change. [results] the cloned silkworm trehalose synthase gene named ApTPS. and the open reading frame of 2487 BP in length, encoding 828 amino acids, egg White predicted molecular weight of 93.19 K D, isoelectric point (P I) 4.61; no signal peptide and transmembrane regions. The protein subcellular localization prediction the protein located in the cytoplasm; protein domain analysis showed that ApTPS has two conserved domains: TPS (22-497 amino acids) and TPP (No. 532-772 amino acids). Tissue specificity analysis showed that ApTPS gene in silkworm larvae in the fat body highest expression; Antheraea pernyi diapause termination process, the amount of increased expression of ApTPS in the fat body and haemolymph, and significantly higher than that of the control group (P0.05), but the expression level in the hemolymph behind the fat body. [Conclusion] the results suggest that ApTPS is involved in carbohydrate metabolism and regulation of tussah pupa diapause played an important role in it, and the pupa diapause termination is closely related.

【作者单位】：沈阳农业大学生物科学技术学院辽宁省昆虫资源工程技术研究中心;辽宁省蚕业科学研究所;
【基金】：现代农业产业技术体系建设专项(CARS-22) 辽宁省农业领域青年科技创新人才培养计划项目(2014010) 辽宁省教育厅一般项目(L2014255,L2015488)
【分类号】：S885.1

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