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
