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脂多糖对家蚕幼虫Toll样受体基因BmToll9-2表达的影响

发布时间:2018-10-29 16:08
【摘要】:【目的】Toll样受体是昆虫Toll信号传导途径的一个重要组成成员,对激活昆虫机体对入侵病原微生物的先天免疫应答起着至关重要的作用。本研究旨在探究脂多糖对家蚕Bombyx mori幼虫中Toll样受体基因BmToll9-2基因表达的影响。【方法】利用RT-PCR分析BmToll9-2基因在家蚕大造品系4龄和5龄幼虫表皮、脂肪体、马氏管、中肠、神经、丝腺、胸部肌肉共7种组织中的表达特征;将脂多糖和大肠杆菌Escherichia coli注射到5龄幼虫体腔内,诱导其免疫系统响应,采用实时荧光定量PCR分析注射后不同时间BmToll9-2基因在中肠中的表达水平。【结果】RT-PCR结果表明,BmToll9-2基因在家蚕4龄幼虫的脂肪体、马氏管、神经和丝腺中高度表达,在5龄幼虫脂肪体、中肠、马氏管和神经中均有较高表达。实时荧光定量PCR结果显示,注射脂多糖能诱导5龄幼虫中肠中BmToll9-2基因转录,在注射后6 h时的诱导效果最好;注射大肠杆菌也能诱导BmToll9-2基因转录,在注射后3和6 h时的诱导效果最好。【结论】家蚕幼虫BmToll9-2基因在脂多糖和大肠杆菌处理后上调表达,提示BmToll9-2受体可能参与昆虫Toll样受体对脂多糖和细菌的识别过程。
[Abstract]:[objective] Toll like receptor is an important member of insect Toll signal transduction pathway, which plays an important role in activating innate immune response of insect body to invading pathogenic microorganisms. The purpose of this study was to investigate the effect of lipopolysaccharide on the expression of BmToll9-2 gene of Toll like receptor gene in silkworm Bombyx mori larvae. [methods] RT-PCR was used to analyze the expression of BmToll9-2 gene in the epidermis, fat body and Markov tube of 4th and 5th instar larvae of silkworm. The expression characteristics of midgut, nerve, silk gland and chest muscle; Lipopolysaccharide (LPS) and Escherichia coli (E. coli) Escherichia coli were injected into the body cavity of 5th instar larva to induce the immune system response. The expression level of BmToll9-2 gene in midgut at different time after injection was analyzed by real-time fluorescence quantitative PCR. [results] RT-PCR results showed that, BmToll9-2 gene was highly expressed in adipose body, Markov duct, nerve and silk gland of 4th instar larvae of Bombyx mori, and was highly expressed in adipose body, midgut, Markov duct and nerve of 5th instar larva. The results of real-time fluorescence quantitative PCR showed that lipopolysaccharide could induce the transcription of BmToll9-2 gene in the midgut of 5th instar larva, and the effect was the best at 6 h after injection. BmToll9-2 gene transcription was also induced by Escherichia coli injection, which was the best at 3 and 6 hours after injection. [conclusion] the expression of BmToll9-2 gene in silkworm larvae was upregulated after lipopolysaccharide and Escherichia coli treatment. These results suggest that BmToll9-2 receptors may be involved in the recognition of lipopolysaccharide and bacteria by insect Toll like receptors.
【作者单位】: 广州大学生命科学学院;
【基金】:国家自然科学基金项目(31501898) 广东省教育厅青年创新人才类项目(2015KQNCX119) 广州市珠江科技新星专项(201710010094) 广州大学新苗计划(201503)
【分类号】:S881.2


本文编号:2298169

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