| 摘要: |
| 【目的】 克隆分析家蚕Bmugt3基因cDNA序列，并对其在家蚕不同组织中的表达进行检测。【方法】 以家蚕ak1和873为供试材料，采用生物信息学方法和RT-PCR技术，对家蚕Bmugt3基因cDNA序列进行克隆和组织表达谱研究，并通过XbaⅠ和Hind Ⅲ双酶切重组质粒，将Bmugt3亚克隆至具有2个反向T7启动子,用于体外诱导合成双链RNA（dsRNA）的L4440载体。【结果】 克隆得到了家蚕Bmugt3基因1 675 bp的cDNA序列，编码462个氨基酸，编码蛋白的分子质量为52.3 ku，等电点6.5；多序列比对显示，Bmugt3基因缺失了C末端的跨膜结构域；聚类分析结果显示，其与家蚕phenol-UGT基因聚合在同一分支上。Bmugt3基因主要在家蚕丝腺中表达。【结论】Bmugt3基因是组织特异性表达基因，可能在家蚕类黄酮素的代谢中起作用。 |
| 关键词: 家蚕 Bmugt3 克隆 组织表达 |
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| 基金项目:陕西省自然科学基金项目（SJ08C108）；安康学院专项科研计划项目（07AKXY026） |
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| Cloning,expression and sequence analysis of cDNA encoding the silkworm Bmugt3 gene |
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KONG Wei qing YANG Jin hong
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| Abstract: |
| 【Objective】 The study was to clone and analyze the cDNA sequence of silkworm Bmugt3 gene and the expression in 5 tissues.【Method】 Using silkworm strain,ak1 and 873,as materials,the cDNA sequence of Bmugt3 gene was cloned by RT-PCR and analyzed by bioinformatics.The gene was subcloned to L4440 vector,which had two T7 polymerase promoter in opposite orientation to synthesize dsRNA in vitro,through double digestion with XbaⅠ and HindⅢ.【Result】 Bmugt3 gene of silkworm with 1 675 bp in length was cloned.426 amino acids were coded and the MW and pI of the predicted protein was 52.3 ku and 6.5 respectively.Bmugt3 had no C-end transmembrane domain by the multiple sequence alignment and it clustered in the same branch with silkworm phenol UGT in the clustering analysis.The Bmugt3 gene expressed mostly in the silk glands in all the tested silkworm tissues.【Conclusion】 The Bmugt3 gene is a tissue specific gene and may function in the metabolism of silkworm flavonoids. |
| Key words: Bombyx mori Bmugt3 cloning tissue expression |
