柞蚕蛹解除滞育过程中海藻糖合成酶基因的表达变化

doi:10.16380/j.kcxb.2016.09.003

摘要/Abstract

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 kD，等电点（pI）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（P<0.05），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。

关键词: 柞蚕, 滞育, 海藻糖合成酶, 基因表达, 滞育解除

Abstract: 【Aim】 This study aims to clone the trehalose-6-phosphate synthase (TPS) gene in Antheraea pernyi, to analyze its tissue distribution, and to profile the expression patterns of the gene during diapause termination so as to provide data support for clarifying the relationship between carbohydrate metabolism and diapause termination in diapause pupae of A. pernyi. 【Methods】 The TPS gene was cloned from the fat body of A. pernyi larvae using PCR and 3′ RACE technology, and the putative amino acid sequence was analyzed by bioinformatics methods. The expression pattern of the gene in different tissues of A. pernyi larvae was analyzed by RT-PCR. The relative expression levels of the gene in haemolymph and fat body in the process of diapause termination of A. pernyi larvae were measured using real-time PCR technology. 【Results】 The TPS gene of A. pernyi was cloned and named ApTPS. It has an open reading frame (ORF) of 2 487 bp, encoding 828 amino acid residues. The encoded protein is estimated to be 93.19 kD with an isoelectric point (pI) about 4.61, and has no signal peptide and transmembrane region. Subcellular localization analysis indicated that ApTPS targets to cytoplasm. Homology search showed that ApTPS has two conserved domains: TPS (22-497 amino acids) and TPP (532-772 amino acids). Tissue specific mRNA expression profiling showed that ApTPS was mainly expressed in the fat body of A. pernyi larvae. In the process of diapause termination, the expression levels of ApTPS in both haemolymph and fat body were up-regulated and significantly higher than those in the control groups (P<0.05). The elevated expression of the gene in the haemolymph, however, occurred later than that in the fat body. 【Conclusion】 The results suggest that ApTPS participates in the regulation of carbohydrate metabolism and plays a significant role in diapause process of A. pernyi pupae. There is a close relationship between ApTPS expression and the diapause termination of A. pernyi pupae.

Key words: Antheraea pernyi, diapause, trehalose-6-phosphate synthase, gene expression, diapause termination

黄伶, 孙良振, 王勇, 汝玉涛, Muhammad IRFAN, 姜义仁, 石生林, 杨瑞生, 李喜升, 秦利. 柞蚕蛹解除滞育过程中海藻糖合成酶基因的表达变化[J]. 昆虫学报, 2016, 59(9): 938-947.

HUANG Ling, SUN Liang-Zhen, WANG Yong, RU Yu-Tao, Muhammad IRFAN, JIANG Yi-Ren, Shi-Sheng-Lin , YANG Rui-Sheng, LI Xi-Sheng, QIN Li. Changes in the expression of trehalose-6-phosphate synthase gene in Antheraea pernyi (Lepidoptera: Saturniidae) during pupal diapause termination[J]. Acta Entomologica Sinica, 2016, 59(9): 938-947.

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柞蚕蛹解除滞育过程中海藻糖合成酶基因的表达变化

Changes in the expression of trehalose-6-phosphate synthase gene in Antheraea pernyi (Lepidoptera: Saturniidae) during pupal diapause termination

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