| 脆性组氨酸三联体基因小干扰RNA的构建及干扰效果检测 |
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| 引用本文： | 林宇翔,于芳,绳纪坡,高宁,马祖兴,胡宝成.脆性组氨酸三联体基因小干扰RNA的构建及干扰效果检测[J].生物技术通讯,2010,21(4):484-487. |
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| 作者姓名： | 林宇翔 于芳 绳纪坡 高宁 马祖兴 胡宝成 |
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| 作者单位： | 1. 军事医学科学院生物工程研究所,北京,100850;福建农林大学,生命科学院,福建,福州,350002
2. 军事医学科学院生物工程研究所,北京,100850 |
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| 基金项目： | 国家自然科学基金，国家重点基础研究发展计划，国家"十一五"支撑项目 |
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| 摘 要： | 目的:构建脆性组氨酸三联体(FHIT)基因的小干扰RNA(siRNA),并检测其对293T细胞内源性FHIT基因表达的干扰效果。方法:根据FHIT基因序列,通过生物信息学网站预测可能的siRNA,从中筛选出2条合理的FHIT-siRNA,将其克隆到siRNA表达载体pSilencer2.1-U6Hygro中,转化大肠杆菌DH5α,挑取阳性克隆进行测序鉴定;将构建成功的FHIT-siRNA重组载体转染人胚肾细胞293T,Western印迹检测siRNA对293T内源性FHIT表达的干扰效果。结果:构建了2个FHIT-siRNA重组质粒,2条siRNA都有干扰作用,其中一条的作用效果可达50%以上。结论:构建的FHIT-siRNA为进一步研究FHIT的功能提供了有利的工具。
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| 关 键 词： | 脆性组氨酸三联体基因 小干扰RNA 表达载体 |
Construction of Small Interfering RNA of Fragile Histidine Tria Gene and Assay of its Interfering Effect |
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| LIN Yu-Xiang,YU Fang,SHENG Ji-Po,GAO Ning,MA Zu-Xing,HU Bao-Cheng.Construction of Small Interfering RNA of Fragile Histidine Tria Gene and Assay of its Interfering Effect[J].Letters in Biotechnology,2010,21(4):484-487. |
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| Authors: | LIN Yu-Xiang YU Fang SHENG Ji-Po GAO Ning MA Zu-Xing HU Bao-Cheng |
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| Institution: | LIN Yu-Xiang1,2,YU Fang1,SHENG Ji-Po1,GAO Ning1,MA Zu-Xing1,HU Bao-Cheng1 1. Beijing Institute of Biotechnology,Beijing 100850,2. School of Life Sciences,Fujian Agriculture and Forestry University,Fuzhou 350002,China |
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| Abstract: | Objective: To construct small interfering RNA(siRNA) of fragile histidine tria(FHIT) gene expression vector, and investigate its interfering effect. Methods: Two FHIT-siRNA were designed and inserted into the pSilencer 2.1-U6 Hygro vector. After confirmation of the constructs by DNA sequencing, human embryo kidney 293T cells were transfected with FHIT-siRNA expression vectors. The interfering effect of FHIT-siRNA was identified by Western blot. Results: Two FHIT-siRNA expression vectors were obtained and co... |
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| Keywords: | fragile histidine tria gene small interfering RNA expression vector |
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