RNF34调控天然免疫的初步研究
发布时间:2018-04-01 11:05
本文选题:RNF 切入点:免疫 出处:《军事医学》2017年01期
【摘要】:目的研究环指蛋白34(RING finger protein 34,RNF34)对天然免疫的调控。方法利用重组PCR方法构建pcDNA3-Flag-RNF34、pcDNA3-Flag-RNF34-ΔFYVE、pcDNA3-Flag-RNF34-ΔCID和pcDNA3-Flag-RNF34-ΔRING质粒,并在HEK293T细胞中瞬时表达;利用双荧光素酶报告基因技术检测RNF34及其3种突变体对仙台病毒(SeV)和N-RIG-Ⅰ激活NF-κB和IFN-β转录活性的影响。结果成功构建了Flag-RNF34及其3种结构域缺失突变体的真核表达质粒;发现在SeV刺激下,RNF34对NF-κB和IFN-β转录活性有明显抑制作用,RNF34-ΔFYVE、RNF34-ΔCID和RNF34-ΔRING与RNF34相比此种抑制作用减弱。同时发现对于N-RIG-Ⅰ激活的NF-κB和IFN-β活性,RNF34及其3种结构域缺失突变体也有相似的抑制作用。结论 RNF34通过负调控RIG-Ⅰ-MAVS信号通路调控天然免疫。
[Abstract]:Objective to study the regulation of innate immunity by cyclic finger protein 34(RING finger protein 34RNF34. methods pcDNA3-Flag-RNF34- 螖 FYVEV pcDNA3-Flag-RNF34- 螖 CID and pcDNA3-Flag-RNF34- 螖 RING were constructed by recombinant PCR method and expressed in HEK293T cells. Double luciferase reporter gene technique was used to detect the effects of RNF34 and its three mutants on the transcriptional activities of Sendai virus (Sendai virus) and N-RIG- 鈪,
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