TNFAIP8干扰质粒的构建及最佳干扰效率质粒的筛选; Construction and identification of interference plasmid targeting on TNFAIP8 | |
Liu WM(刘文明) ; Yang JJ(杨晶晶) ; Hu RY(胡如意) ; Qiu XF(邱兴烽) ; Shi CY(石春燕) ; Qi ZQ(齐忠权) ; Liu ZC(刘忠臣) ; Zhuang GH(庄国洪) | |
2015-05-20 | |
关键词 | TNFAIP8 RNA干扰 重组质粒 筛选 TNFAIP8 RNA interference Recombinant plasmid Screen |
英文摘要 | 目的:构建并筛选出干扰效率最佳的TnfAIP8-SHrnA-P SIrEn-rETrO Q干扰质粒。方法:通过生物软件选择3个TnfAIP8基因干扰位点,构建干扰质粒并测序验证,将干扰质粒及对照质粒分别转染至A549细胞,通过rT-PCr、WESTErn blOT检测干扰效率。结果:经rT-PCr和WESTErn-blOT证实TnfAIP8-SHrnA-P SIrEn-rETrO Q干扰质粒能有效干扰并抑制细胞内TnfAIP8基因的表达,通过流式检测发现降低TnVAIP8表达可以提高细胞对A dr5SC fV诱导凋亡的敏感性。结论:成功构建和设计了对TnfAIP8基因具有显著干扰效率的干扰质粒,为进一步研究TnfAIP8基因的功能奠定了基础。; Objective: To construct and screen the high efficiency interference plasmid of TFAIP8-shRNA-p SIRENRetro Q.Methods: Selected and synthesized three Target Sequence of TNFAIP8 shRNA1,TNFAIP8 shRNA2,TNFAIP8 shRNA3,and construct the TNFAIP8 interference plasmid.Transfection TNFAIP8-shRNA-p SIREN-Retro Q interference plasmid to A549 cells.Filter out the highest interference efficiency plasmid by detecting the mRNA and protein levels using RT-PCR and Western blot methods.Results: We successfully design and built three TNFAIP8-shRNA-p SIREN-Retro Q interference plasmids,and screen out the highest efficiency interference plasmid.Conclusion: Three interference plasmids targeting the TNFAIP8 gene have been constructed successfully and provide a useful tool for studying the function of TNFAIP8.; 国家自然科学基金项目(81272720); 福建省卫计委医学创新课题(2014-CXB-43); 厦门市科技计划项目(3502Z2083008)资助 |
语种 | zh_CN |
内容类型 | 期刊论文 |
源URL | [http://dspace.xmu.edu.cn/handle/2288/123776] ![]() |
专题 | 医学院-已发表论文 |
推荐引用方式 GB/T 7714 | 刘文明,杨晶晶,胡如意,等. TNFAIP8干扰质粒的构建及最佳干扰效率质粒的筛选, Construction and identification of interference plasmid targeting on TNFAIP8[J],2015. |
APA | 刘文明.,杨晶晶.,胡如意.,邱兴烽.,石春燕.,...&庄国洪.(2015).TNFAIP8干扰质粒的构建及最佳干扰效率质粒的筛选.. |
MLA | 刘文明,et al."TNFAIP8干扰质粒的构建及最佳干扰效率质粒的筛选".(2015). |
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