HMGN2基因干扰质粒构建及干扰效率测定 |
| |
引用本文: | 赵静,邓璐霞,韩琴,吴桂霞,曹玥,李夏,范波,陈善泽,高祥,黄宁. HMGN2基因干扰质粒构建及干扰效率测定[J]. 四川生理科学杂志, 2010, 32(2): 54-56 |
| |
作者姓名: | 赵静 邓璐霞 韩琴 吴桂霞 曹玥 李夏 范波 陈善泽 高祥 黄宁 |
| |
作者单位: | 1. 四川大学华西医学中心感染与免疫实验室,四川,成都,610041;成都中医药大学基础医学院,四川,成都,610000 2. 四川大学华西医学中心感染与免疫实验室,四川,成都,610041 |
| |
基金项目: | 国家自然科学基金,CMB基金 |
| |
摘 要: | 目的:构建HMGN2基因的干扰质粒,为深入研究HMGN2基因在信号通路中的作用提供有效手段。方法:根据文献选择两个HMGN2基因的干扰位点,合成两个干扰片段定向克隆到psilencer-4.1的干扰载体并测序验证。将干扰质粒转染至A549细胞,并通过检测RT-PCR产物量以获得干扰效率。结果:RT—PCR产物检测结果显示干扰质粒psilencer-4.1-HMGN2—2干扰效率最高,其HMGN2表达量降低了70%左右。结论:成功构建了对HMGN2基因具有显著干扰效率的psilencer-4.1-HMGN2干扰质粒,为进一步研究HMGN2基因的功能打下了基础。
|
关 键 词: | RNA干扰 干扰质粒RT-PCR HMGN2基因 |
Construction and identification of interference plasmid of HMGN2 |
| |
Affiliation: | Zhao Jing, Deng Lu-xia, Han Qin, et al (1. Research Unit of Infection and Immunity, West China School of Preclinical and Forensic Medicine, Sichuan University, Sichuan Chengdu 610041;2. Pathological Department of Chengdu University of Chinese Medicine, Sichuan Chengdu 610041) |
| |
Abstract: | Objective: To construct highly efficient interference plasmid aiming at HMGN2 gene, and provide an effective way for studying the function of HMGN2 gene. Methods:Two interference sites of HMGN2 gene were selected from documents and two interference fragments and one negative control fragment were synthesized, then they were cloned into the psilencer-cmv-4. 1 plasmid. After sequencing, the plasmids were transfected into A549 cells. The interference efficiency of the plasmid on the target gene was detected by semiquantitative RT-PCR. Results: The level of HMGN2 mRNA in A549 ceils contained interference plasmids decreased by 70% compared with negative control plasmid. Conclusion: The effective interference plasmid against HMGN2 gene had been constructed and identified successfully, which may provide a basis for studying the function of HMGN2 gene in signal transduetion pathway. |
| |
Keywords: | RNA interference Interference plasmid RT-PCR HMGN2 |
本文献已被 维普 万方数据 等数据库收录! |
|