【摘要】：【目的】Toll樣受體是昆蟲Toll信號傳導(dǎo)途徑的一個重要組成成員,對激活昆蟲機體對入侵病原微生物的先天免疫應(yīng)答起著至關(guān)重要的作用。本研究旨在探究脂多糖對家蠶Bombyx mori幼蟲中Toll樣受體基因BmToll9-2基因表達的影響。【方法】利用RT-PCR分析BmToll9-2基因在家蠶大造品系4齡和5齡幼蟲表皮、脂肪體、馬氏管、中腸、神經(jīng)、絲腺、胸部肌肉共7種組織中的表達特征;將脂多糖和大腸桿菌Escherichia coli注射到5齡幼蟲體腔內(nèi),誘導(dǎo)其免疫系統(tǒng)響應(yīng),采用實時熒光定量PCR分析注射后不同時間BmToll9-2基因在中腸中的表達水平�！窘Y(jié)果】RT-PCR結(jié)果表明,BmToll9-2基因在家蠶4齡幼蟲的脂肪體、馬氏管、神經(jīng)和絲腺中高度表達,在5齡幼蟲脂肪體、中腸、馬氏管和神經(jīng)中均有較高表達。實時熒光定量PCR結(jié)果顯示,注射脂多糖能誘導(dǎo)5齡幼蟲中腸中BmToll9-2基因轉(zhuǎn)錄,在注射后6 h時的誘導(dǎo)效果最好;注射大腸桿菌也能誘導(dǎo)BmToll9-2基因轉(zhuǎn)錄,在注射后3和6 h時的誘導(dǎo)效果最好�！窘Y(jié)論】家蠶幼蟲BmToll9-2基因在脂多糖和大腸桿菌處理后上調(diào)表達,提示BmToll9-2受體可能參與昆蟲Toll樣受體對脂多糖和細菌的識別過程。
[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.
【作者單位】： 廣州大學(xué)生命科學(xué)學(xué)院;
【基金】：國家自然科學(xué)基金項目(31501898) 廣東省教育廳青年創(chuàng)新人才類項目(2015KQNCX119) 廣州市珠江科技新星專項(201710010094) 廣州大學(xué)新苗計劃(201503)
【分類號】：S881.2
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本文編號：2298169