| 人miR-34a靶基因的生物信息学分析及载体构建 |
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| 引用本文： | 杨聚荣,蔡广研,洪权,谢院生,傅博,吕杨,刘晓峦,崔敬,陈香关.人miR-34a靶基因的生物信息学分析及载体构建[J].军医进修学院学报,2010,31(5):467-469. |
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| 作者姓名： | 杨聚荣 蔡广研 洪权 谢院生 傅博 吕杨 刘晓峦 崔敬 陈香关 |
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| 作者单位： | 解放军总医院肾科全军肾脏病研究所,北京,100853 |
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| 基金项目： | 国家“973”重点基础研究发展规划项目(2007CB507403)~~ |
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| 摘 要： | 目的对人miR-34a（hsa-miR-34a）下游靶基因进行预测,并构建含自噬相关基因靶结合序列的GFP报告基因载体。方法使用TargetScan Release 5.1和RNA22软件分析hsa-miR-34a可能的靶基因,并利用Gominer软件对靶基因功能进行分析。根据预测结果,合成含自噬相关基因hsa-miR-34a靶序列的寡核苷酸,以pEGFPC2为载体构建系列质粒,并检测各载体的真核表达情况。结果预测结果显示hsa-miR-34a共有2904个下游靶基因,功能涵盖生物过程、细胞组分、分子功能三大类,其中与自噬作用直接相关的靶基因有beclin 1和sestrin 2。构建含beclin 1和sestrin 2作用位点的GFP载体共5个,质粒转染Hela细胞后在荧光显微镜下可见绿色荧光。结论成功构建hsa-miR-34a下游自噬相关基因靶位点序列GFP报告基因载体,为进一步研究hsa-miR-34a对自噬的调控作用奠定基础。
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| 关 键 词： | hsa-miR-34a 自噬 基因 报告 |
Bioinformatics analysis of hsa-miR-34a target genes and construction of green fluorescence protein reporter gene vectors |
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| YANG Ju-rong,CAI Guang-yan,HONG Quan,XIE Yuan-sheng,FU Bo,LV Yang,LIU Xiao-luan,CUI Jing,CHEN Xiang-mei.Bioinformatics analysis of hsa-miR-34a target genes and construction of green fluorescence protein reporter gene vectors[J].Academic Journal of Pla Postgraduate Medical School,2010,31(5):467-469. |
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| Authors: | YANG Ju-rong CAI Guang-yan HONG Quan XIE Yuan-sheng FU Bo LV Yang LIU Xiao-luan CUI Jing CHEN Xiang-mei |
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| Institution: | YANG Ju-rong,CAI Guang-yan,HONG Quan,XIE Yuan-sheng,FU Bo,LV Yang,LIU Xiao-luan,CUI Jing,CHEN Xiang-mei Department of Nephrology,Chinese PLA Center of Kidney Diseases,Chinese PLA General Hospital,Beijing 100853,China |
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| Abstract: | Objective To predict the hsa-miR-34a target genes and construct green fluorescence protein(GFP) reporter gene vectors containing autophagy-associated target genes.Methods Biological targets of hsa-miR-34a were predicted using the online program TargetScan Release 5.1 and RNA22.Their molecular function was analyzed using software Gominer.Oligonucleotide sequences of hsa-miR-34a binding sites were synthesized and cloned into pEGFPC2 after digestion with EcoR I and BamH I.Recombinant plasmids was confirmed by ... |
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| Keywords: | hsa-miR-34a Autophagy Genes Reporter |
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