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| 人HMGN5基因shRNA慢病毒载体构建与RNAi效率的鉴定 | |
| 引用本文： | 陈鹏,马忠森,王秀丽,李博,任锦,胡娱新,郏博,刘锋,石莹.人HMGN5基因shRNA慢病毒载体构建与RNAi效率的鉴定[J].中国实验诊断学,2011,15(1):7-10. |
| 作者姓名： | 陈鹏 马忠森 王秀丽 李博 任锦 胡娱新 郏博 刘锋 石莹 |
| 作者单位： | 1. 吉林大学第二医院,吉林,长春,130041 2. 吉林大学第二医院,吉林,长春,130041;吉林省职业病防治院 3. 吉林大学白求恩医学院临床二系 4. 日本千叶大学融合科学研究所 5. 东北师范大学遗传与细胞研究所 |
| 基金项目： | 国家自然科学基金资助课题 |
| 摘 要： | 目的构建人HMGN5基因的shRNA慢病毒载体并鉴定在肺癌A549和H1299细胞上的沉默效率。方法设计HMGN5基因特异性siRNA靶点,构建于慢病毒pLL-3.7载体,并筛选获得有效的shRNA慢病毒载体,在293T细胞包装成病毒颗粒,将其感染肺癌A549和H1299细胞,应用Real-time PCR方法从mRNA水平上检测HMGN5的沉默效率。结果构建的慢病毒载体shRNA的PCR鉴定和测序正确,包装病毒后滴度达到5×108TU/ml。shRNA慢病毒颗粒感染A549和H1299细胞后HMGN5基因的mRNA表达量较阴性对照载体慢病毒感染组分别下降了71.7%和50.7%。结论成功构建了HMGN5基因的shRNA慢病毒表达载体,在分子水平能够有效沉默靶基因,为探讨HMGN5在肿瘤基因治疗中的作用奠定了基础。 |
| 关 键 词： | HMGN5 shRNA 慢病毒 |
Construction of shRNA lentiviral vector targeting human HMGN5gene and identification of RNAi efficiency |
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| CHEN Peng,MA Zhong-sen,WANG Xiu-li,et al..Construction of shRNA lentiviral vector targeting human HMGN5gene and identification of RNAi efficiency[J].Chinese Journal of Laboratory Diagnosis,2011,15(1):7-10. | |
| Authors: | CHEN Peng MA Zhong-sen WANG Xiu-li |
| Institution: | CHEN Peng,MA Zhong-sen,WANG Xiu-li,et al.(Second Hospital,Jilin University,Changchun 130041,China) |
| Abstract: | Objective To construct shRNA lentiviral vectors targeting humanHMGN5 gene and detect its effect of gene silence in A549 and H1299 cells.Methods The specific siRNA sequences targeting human HMGN5 gene were cloned into pLL-3.7 lentiviral vector.The lentivirus particles were packaged and HMGN5 specific shRNA was transmitted into A549 and H1299 cells after screeningfor the valid siRNA.HMGN5 efficiency of silencing was determined by Real-time PCR from mRNA level.Results It revealed shRNA plasmids was correctly c... |
| Keywords: | HMGN5 shRNA |
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