| 摘要: |
| 【目的】对家蚕(Bombyx mori)糖原合成酶激酶3α(GSK3α)蛋白的氨基酸序列进行分析，并检测家蚕不同发育时期及蜕皮激素、饥饿处理下，GSK3α基因mRNA转录水平的变化。【方法】以家蚕为供试材料，采用RT PCR和荧光定量PCR方法，研究家蚕GSK3α在不同发育期的表达和转录情况，及激素和饥饿处理对其转录的影响。【结果】家蚕GSK3α基因由2个外显子和1个内含子构成，其中内含子长度为1 411 bp，预测家蚕GSK3α蛋白质分子质量为33.78 ku，等电点为7.61。氨基酸系统进化树和序列多重联配分析结果表明，家蚕GSK3α与所选几种昆虫GSK3α的序列一致性较高(79%～91%)。家蚕GSK3α mRNA在幼虫取食期转录水平较高，在蜕皮期转录水平较低。在注射后6 h内，蜕皮激素对家蚕GSK3α基因mRNA的转录无诱导作用，但在注射12～24 h有较强的诱导作用，而且在注射12 h时诱导活性达到最高，之后逐渐降低。饥饿处理对家蚕GSK3α mRNA的转录水平影响较小。【结论】家蚕GSK3α在昆虫的进化过程中是比较保守的，其表达受到蜕皮激素的调控，饥饿处理对其转录水平影响较小。 |
| 关键词: 家蚕 糖原合成酶激酶 序列分析 |
| DOI： |
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| 基金项目:河南省教育厅自然科学研究计划项目（2011B180021） |
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| Amino acid sequence analysis and study on mRNA transcription level of Bombyx mori glycogen synthase kinase 3 α |
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| Abstract: |
| 【Objective】The amino acid sequence of Bombyx mori glycogen synthase kinase 3α (BmGSK3α) was analyzed in this study.The mRNA transcription level of BmGSK3α was further stuied in different developed stages and after molting hormone and starvation treatments.【Method】Temporal expression profiles of BmGSK3α,effects of hormone and starvation treatment on BmGSK3α transcript were investigated by using Real-time PCR methods.【Result】The sequence analysis of BmGSK3α found that BmGSK3α gene consists of 2 exons and one intron of 1 411 bp in size.The predicted molecular weight and pI of BmGSK3α are 33.78 ku and 7.61 respectively.Amino acid sequence alignments indicated that BmGSK3α shared high identity with other reported GSK proteins (from 79% to 91%).The expression level of BmGSK3α increased in larva feeding stage and decreased in molting stage.BmGSK3α transcript had no significant difference after 6 h treatment of 20E and increased after 12 h of 20E treatment then decreased gradually.Further starvation treatment found that starvation had limited influence on BmGSK3α expression.【Conclusion】BmGSK3α is conserved in evolution process of insects.The expression level of BmGSK3α is regulated by 20E and starvation has limited influence on BmGSK3α expression. |
| Key words: Bombyx mori glycogen synthase kinase sequence analysis |
