| Stathmin SiRNA表达载体抑制stathmin的表达 |
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| 引用本文： | 段佳,黄岚珍,何晓松,易世江,范才文,肖胜军,雷迅.Stathmin SiRNA表达载体抑制stathmin的表达[J].生物技术,2011,21(4). |
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| 作者姓名： | 段佳 黄岚珍 何晓松 易世江 范才文 肖胜军 雷迅 |
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| 作者单位： | 1. 桂林医学院附属医院耳鼻喉头颈外科,广西桂林,541001
2. 桂林医学院科学实验中心,广西桂林,541004 |
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| 基金项目： | 广西科学研究与技术开发课题，广西卫生厅重点科研课题，广西科技厅项目 |
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| 摘 要： | 目的:构建stathmin特异性SiRNA质粒表达载体,探讨其对鼻咽癌5-8F细胞stathmin的沉默作用.方法:合成用于stathmin基因特异性干扰表达的DNA片段,经退火形成双链DNA片段,片段克隆到质粒表达载体pGenesil 1.1上.载体导入JM109菌株进行筛选与扩增,采用酶切和测序对克隆表达载体进行鉴定.应用脂质体将鉴定后的重组表达质粒载体转入鼻咽癌5 -8F细胞,RT-PCR与Western Blot分析stathmin基因表达.结果:经酶切和测序鉴定,插入SiRNA质粒表达载体的stathmin特异性碱基序列和方向正确.重组质粒表达载体转染鼻咽癌细胞后,细胞转染效率达78.8 ±6.8％,stathmin基因在鼻咽癌中的表达明显下降.结论:构建的stathmin基因SiRNA质粒表达载体能抑制stathmin的表达.
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| 关 键 词： | stathmin RNA干扰 质粒 基因治疗 |
Stathmin SiRNA Plasmid Expression Vector Suppressing Expression of Stathmin |
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| DUAN Jia,HUANG Lan-zhen,HE Xiao-song,YI Shi-jiang,FAN Cai-wen,XIAO Sheng-jun,LEI Xun.Stathmin SiRNA Plasmid Expression Vector Suppressing Expression of Stathmin[J].Biotechnology,2011,21(4). |
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| Authors: | DUAN Jia HUANG Lan-zhen HE Xiao-song YI Shi-jiang FAN Cai-wen XIAO Sheng-jun LEI Xun |
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| Institution: | DUAN Jia1,HUANG Lan-zhen2,HE Xiao-song1,YI Shi-jiang1,FAN Cai-wen2,XIAO Sheng-jun2,LEI Xun1*(1.Department of Otorhinolaryngology,The Afiliated Hospital of Guilin Medical College,Guilin 541001,China,2.Center Laboratory of Guilin Medical College,Guilin 541004,China) |
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| Abstract: | Objective:Constructing stathmin specific SiRNA plasmid expression vector and investigating the silence effect of SiRNA plasmid expression vector on stathmin in 5-8F cell line.Method:Synthesize stathmin specific interfering expression DNA fragments,obtain doubled DNA fragment by annealing and combine the fragment with plasmid pGenesil1.1.The recombinant vectors were transferred and screened in JM109 strains.Then,the recombinant vectors were amplified and identified by di-enzyme digestion and DNA sequencing.T... |
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| Keywords: | stathmin RNAi plasmid gene therapy |
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