家蠶Met1基因的表達(dá)與組織定位分析
發(fā)布時(shí)間：2018-10-19 20:26
【摘要】：選擇家蠶基因組中與果蠅Dm Met(Methoprene tolerant protein)同源的基因Bm Met1進(jìn)行研究。預(yù)測(cè)Dm Met和Bm Met1的結(jié)構(gòu)域均是一個(gè)b HLH-PAS(basic helix-loop-helix Per-ARNT-SIM)型轉(zhuǎn)錄因子,具有典型的DNA結(jié)合結(jié)構(gòu)域。在原核系統(tǒng)表達(dá)獲得了Bm Met1蛋白并制備了抗體。qRT-PCR和Western blotting檢測(cè)結(jié)果顯示,無(wú)論是Bm Met1基因mRNA的轉(zhuǎn)錄還是Bm Met1蛋白的表達(dá),在家蠶5齡中后期和吐絲期的翅原基組織中都呈現(xiàn)較高水平,而在蛹期呈現(xiàn)較低的水平。通過(guò)免疫組織化學(xué)方法分析Bm Met1的組織定位,結(jié)果顯示該蛋白質(zhì)在5齡第6天和吐絲期家蠶胸節(jié)表皮、脂肪體及翅原基等組織中都有較高水平的表達(dá),在蛹期上述部位和組織的表達(dá)水平較低。分別將蛻皮激素(20E)和保幼激素類似物(Methoprene)按照2μg/頭的劑量注射到5齡第3天幼蟲第2~3胸節(jié)間,qRT-PCR檢測(cè)幼蟲翅原基組織中的Bm Met1受到Methoprene的誘導(dǎo)上調(diào)表達(dá),且在處理2 h后表達(dá)水平最高。上述結(jié)果暗示Bm Met1可能參與了保幼激素對(duì)家蠶翅原基生長(zhǎng)分化的調(diào)控。
[Abstract]:The Bm Met1 gene homologous to Dm Met (Methoprene tolerant protein) of Drosophila melanogaster was selected from the genome of Bombyx mori (Bombyx mori). The predicted domains of both Dm Met and Bm Met1 are b HLH-PAS (basic helix-loop-helix Per-ARNT-SIM) transcription factors with typical DNA binding domains. Bm Met1 protein was expressed in prokaryotic system and antibody was prepared. The results of qRT-PCR and Western blotting showed that both the transcription of Bm Met1 gene mRNA and the expression of Bm Met1 protein were detected. The level of wing primordium of silkworm was higher in the middle and later stage of silkworm and silkworm at silkworm than in pupal stage. The localization of Bm Met1 was analyzed by immunohistochemical method. The results showed that the protein was highly expressed in the epidermis, adipose body and wing primordium of silkworm on the 6th day of 5th instar and in silkworm at the stage of silkworm (Bombyx mori). The expression level of the above mentioned parts and tissues was low in pupa stage. 2 渭 g / head (Methoprene) was injected into the second thoracic internode of the 3rd instar larva on the 3rd day of 5th instar respectively. QRT-PCR was used to detect the up-regulated expression of Bm Met1 in the larval wing primordium tissue induced by Methoprene. The expression level was the highest after 2 h treatment. These results suggest that Bm Met1 may be involved in the regulation of juvenile hormone on the growth and differentiation of wing primordium of Bombyx mori.
【作者單位】： 華南師范大學(xué)生命科學(xué)學(xué)院 廣州市昆蟲發(fā)育調(diào)控與應(yīng)用重點(diǎn)實(shí)驗(yàn)室;
【基金】：國(guó)家自然科學(xué)基金重點(diǎn)項(xiàng)目(No.31330071)
【分類號(hào)】：S881.2
本文編號(hào)：2282269
[Abstract]:The Bm Met1 gene homologous to Dm Met (Methoprene tolerant protein) of Drosophila melanogaster was selected from the genome of Bombyx mori (Bombyx mori). The predicted domains of both Dm Met and Bm Met1 are b HLH-PAS (basic helix-loop-helix Per-ARNT-SIM) transcription factors with typical DNA binding domains. Bm Met1 protein was expressed in prokaryotic system and antibody was prepared. The results of qRT-PCR and Western blotting showed that both the transcription of Bm Met1 gene mRNA and the expression of Bm Met1 protein were detected. The level of wing primordium of silkworm was higher in the middle and later stage of silkworm and silkworm at silkworm than in pupal stage. The localization of Bm Met1 was analyzed by immunohistochemical method. The results showed that the protein was highly expressed in the epidermis, adipose body and wing primordium of silkworm on the 6th day of 5th instar and in silkworm at the stage of silkworm (Bombyx mori). The expression level of the above mentioned parts and tissues was low in pupa stage. 2 渭 g / head (Methoprene) was injected into the second thoracic internode of the 3rd instar larva on the 3rd day of 5th instar respectively. QRT-PCR was used to detect the up-regulated expression of Bm Met1 in the larval wing primordium tissue induced by Methoprene. The expression level was the highest after 2 h treatment. These results suggest that Bm Met1 may be involved in the regulation of juvenile hormone on the growth and differentiation of wing primordium of Bombyx mori.
【作者單位】： 華南師范大學(xué)生命科學(xué)學(xué)院 廣州市昆蟲發(fā)育調(diào)控與應(yīng)用重點(diǎn)實(shí)驗(yàn)室;
【基金】：國(guó)家自然科學(xué)基金重點(diǎn)項(xiàng)目(No.31330071)
【分類號(hào)】：S881.2
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