针对DREAM基因序列外显子的siRNA设计  

Design of siRNA focusing on the exons of DREAM gene

作　　者：张永福 项红兵[2] 毕小宝 劳建新 姚侠 程秋菊 

机构地区：[1]广州市妇女儿童医疗中心麻醉科,广东广州510180 [2]广东省中医院麻醉科,广东广州510180

出　　处：《现代生物医学进展》2012年第1期42-45,共4页Progress in Modern Biomedicine

基　　金：广州市医药卫生科技项目(2009-YB-063)

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。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.

关 键 词：DREAM 外显子 SIRNA 

分 类 号：R-058[医药卫生] Q75[生物学—分子生物学]