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| RNAi技术干扰人喉癌Hep-2细胞PTTG1基因的初步研究 | |
| 引用本文： | 王云龙,孙新城,徐桂超,昌静峰,李玉林,王国强,刘旺根,董彩文.RNAi技术干扰人喉癌Hep-2细胞PTTG1基因的初步研究[J].实用医学杂志,2011,27(16):2913-2917. |
| 作者姓名： | 王云龙 孙新城 徐桂超 昌静峰 李玉林 王国强 刘旺根 董彩文 |
| 作者单位： | 1. 郑州职业技术学院,450121 2. 郑州大学,河南省新乡市,453007 3. 河南省生物工程技术研究中心,郑州市,450001 |
| 基金项目： | 河南省杰出人才创新基金 |
| 摘 要： | 目的:探索对人PTTG1基因mRNA序列有较高干扰效率的位点,构建对PTTG1基因的表达有较高抑制效率的重组质粒。方法:筛选、合成1对互补DNA单链,退火为双链后与双酶切后的质粒载体连接,构建了表达短发夹RNA的1种重组质粒plk0.1-puro/PTTG1。重组质粒以不同浓度转染,并于转染后不同时间收集细胞,以β-actin为内参,采用RT-PCR和Western blot技术分别检测PTTG1基因的mRNA、蛋白质的相对表达水平。结果:与空白对照组相比,6孔板中每孔转染质粒plk0.1-puro/PTTG 11000ng以上、转染超过12d时,PTTG1基因mRNA、蛋白质的相对表达水平下降70%以上。结论:重组质粒plk0.1-puro/PTTG1,在6孔细胞培养板中以1000ng/孔转染12d以上时,有效抑制了人PTTG1基因的表达,在关于PTTG1基因的研究中具有较好的应用价值。 |
| 关 键 词： | RNAi PTTG1基因 shRNA |
The initial research On the inhibition effects of PTTG1 gene by RNA interference in laryngeal carcinoma Hep-2 cells |
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| WANG Yun-long,SUN Xin-cheng,XU Gui-chao,CHANG Jing-feng,LI Yu-lin,WANG Guo-qiang,LIU Wang-gen,DONG Cai-wen.The initial research On the inhibition effects of PTTG1 gene by RNA interference in laryngeal carcinoma Hep-2 cells[J].The Journal of Practical Medicine,2011,27(16):2913-2917. | |
| Authors: | WANG Yun-long SUN Xin-cheng XU Gui-chao CHANG Jing-feng LI Yu-lin WANG Guo-qiang LIU Wang-gen DONG Cai-wen |
| Institution: | WANG Yun-long*,SUN Xin-cheng,XU Gui-chao,CHANG Jing-feng,LI Yu-lin,WANG Guo-qiang,LIU Wang-gen,DONG Cai-wen.*Zhengzhou Technical College,Zhengzhou 450121,China [ |
| Abstract: | Objective To explore the interference sites with high efficiency for the mRNA of human PTTG1 gene,and build on the recombinant plasmids which can restrain the expression of PTTG1 gene.Methods Two pairs of single-stranded DNA sequences were selected and synthesised,and double-stranded DNA molecules were obtained after annealing.The two molecules were linked respectively to the vector plk0.1-puro plasmid which was digested,then two kinds of recombinant plasmids named plk0.1-puro/PTTG1 was successfully constru... |
| Keywords: | RNAi PTTG1 gene shRNA |
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