家蚕Met1基因的表达与组织定位分析
发布时间：2018-10-19 20:26
【摘要】：选择家蚕基因组中与果蝇Dm Met(Methoprene tolerant protein)同源的基因Bm Met1进行研究。预测Dm Met和Bm Met1的结构域均是一个b HLH-PAS(basic helix-loop-helix Per-ARNT-SIM)型转录因子,具有典型的DNA结合结构域。在原核系统表达获得了Bm Met1蛋白并制备了抗体。qRT-PCR和Western blotting检测结果显示,无论是Bm Met1基因mRNA的转录还是Bm Met1蛋白的表达,在家蚕5龄中后期和吐丝期的翅原基组织中都呈现较高水平,而在蛹期呈现较低的水平。通过免疫组织化学方法分析Bm Met1的组织定位,结果显示该蛋白质在5龄第6天和吐丝期家蚕胸节表皮、脂肪体及翅原基等组织中都有较高水平的表达,在蛹期上述部位和组织的表达水平较低。分别将蜕皮激素(20E)和保幼激素类似物(Methoprene)按照2μg/头的剂量注射到5龄第3天幼虫第2~3胸节间,qRT-PCR检测幼虫翅原基组织中的Bm Met1受到Methoprene的诱导上调表达,且在处理2 h后表达水平最高。上述结果暗示Bm Met1可能参与了保幼激素对家蚕翅原基生长分化的调控。
[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.
【作者单位】： 华南师范大学生命科学学院 广州市昆虫发育调控与应用重点实验室;
【基金】：国家自然科学基金重点项目(No.31330071)
【分类号】：S881.2
本文编号：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.
【作者单位】： 华南师范大学生命科学学院 广州市昆虫发育调控与应用重点实验室;
【基金】：国家自然科学基金重点项目(No.31330071)
【分类号】：S881.2
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