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海藻糖酶是海藻糖代谢过程中的一个关键酶,在昆虫发育和能量调节中具有重要作用,为进一步探讨海藻糖酶基因的功能,本文分析了飞蝗Locusta migratoria一可溶型海藻糖酶基因的氨基酸序列并对其mRNA表达特性进行了研究。结果表明:该海藻糖酶(TRE,GenBank登录号:FJ795020)不含有跨膜结构。系统进化树分析结果显示,该酶与大豆蚜Aphis glycines、豌豆蚜Acyrthosiphon pisum、褐飞虱Nilaparvata lugens和灰飞虱Laodelphax striatella可溶型海藻糖酶具有较近的亲缘关系,因此我们将该酶的基因命名为LmTre-1。对该基因在不同组织和发育时期表达量的荧光定量PCR分析表明:LmTre-1在卵发育前期、中期的表达量都很低,卵发育后期表达量显著提高;LmTre-1在5龄若虫和成虫被检测的组织部位中均有表达,在体壁中的表达量最高,其次是在脂肪体、肌肉、气管、精巢及卵巢中;5龄飞蝗刚蜕皮后LmTre-1在体壁中的表达量较高,随着生长发育其表达量逐渐降低;LmTre-1在成虫发育期体壁中稳定高表达。LmTre-1的mRNA表达特性与几丁质合成酶1基因非常相似,据此推测该基因可能与体壁几丁质的合成相关。本研究为深入探讨该基因的生理功能提供了重要的基础数据,并为以海藻糖酶为杀虫靶标的农药筛选奠定实验基础。
Trehalase is a key enzyme in trehalose metabolism and plays an important role in insect development and energy regulation. In order to further explore the function of trehalase gene, trehalosease was analyzed in this paper. Locusta migratoria a soluble trehalase gene The amino acid sequence and its mRNA expression characteristics were studied. The results showed that the trehalase (TRE, GenBank accession number: FJ795020) does not contain a transmembrane structure. The results of phylogenetic tree analysis showed that the enzyme has a close genetic relationship with Aphis glycines, Acyrthosiphon pisum, Nilaparvata lugens and Laodelphax striatella soluble trehalase, so we named the gene of the enzyme For LmTre-1. Quantitative real-time PCR analysis of the expression level of LmTre-1 in different tissues and developmental stages showed that the expression level of LmTre-1 in the early and mid-stage of egg development was very low and the expression of LmTre- Adults were detected in the tissue part of the expression, the highest expression in the body wall, followed by the fat body, muscle, trachea, testis and ovary; locust just after 5 days LmTre-1 molting in the body wall The expression of LmTre-1 was stable and high in the body wall of adult during development. LmTre-1 mRNA expression characteristics and chitin synthase 1 gene is very similar, suggesting that the gene may be related to the synthesis of the body wall chitin. This study provided important basic data for in-depth exploration of the physiological functions of this gene and laid the experimental foundation for the screening of pesticides based on trehalase.