| 张永福1 项红兵2 毕小宝1 劳建新1 姚侠1 程秋菊1.针对DREAM 基因序列外显子的siRNA 设计[J].,2012,12(1):42-45 |
| 针对DREAM 基因序列外显子的siRNA 设计 |
| Design of siRNA focusing on the exons of DREAM gene* |
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| DOI： |
| 中文关键词: DREAM 外显子 siRNA |
| 英文关键词: DREAM Exons Small interfering RNA |
| 基金项目: |
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| 中文摘要: |
| 目的：针对DREAM 基因中外显子序列，设计并筛选出起作用的siRNA，为进一步研究以DREAM 为靶标的基因治疗提供
依据。方法：通过利用生物信息学的方法设计出66 条潜在的针对DREAM 基因中外显子序列的siRNA 序列。潜在的序列根据
G+C 含量分析及Genbank BLAST 检测，我们筛选了三条较理想的DREAM 基因siRNA 靶标，并将其构建到pENTR/H1/TO 载体
中，转染细胞后提取总蛋白并用Western blot 方法检测DREAM 蛋白的表达水平。结果：示外显子9 中5' 末端位置为1253 的序列
能够抑制80%的DREAM 的蛋白表达。结论：这一研究结果为进一步siRNA 类药物的实验研究提供了理论基础。 |
| 英文摘要: |
| Objective: According to the sequence of exons in the DREAM gene, functional siRNA was designed and screened.
This provides a theoretical basis on the further study of DREAM-targeted gene therapy. Methods: Focusing on the exons of DREAM
gene respectively, 66 siRNA candidate targets were obtained following bioinformatics methods. Three ideal siRNA targets were further
screened according to the contents of G+C and Genbank BLAST analysis, and they were constructed into the pENTR/H1/TO vector, the
constructs were then transfected into cells, the total proteins were extracted and to detect the expression of DREAM protein using Western
blot analysis. Results: The sequence located in exon 9 whose 5' end location is 1253 can inhibit 80% expression of DREAM protein.
Conclusion: It would lay a foundation for the further experimental researches on the siRNA-like drug design for the DREAM. |
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