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IFN-β基因启动子调控区域的分析及IRF-3报告基因的构建 被引量：1

Analysis of regulatory elements of IFN-β gene promoter and construction of IRF-3 reporter gene
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摘要 目的对IRF-3的主要靶基因———小鼠IFN-β基因启动子区域进行分析并构建IRF-3报告基因载体。方法将小鼠IFN-β基因启动子全序列以及不同截短片段克隆到无启动子的pGF3basic/enhancer质粒中,以EGFP为报告基因,转染小鼠Raw264.7巨噬细胞系后观察细胞对LPS刺激的反应。结果IFN-β启动子PRD调控区域上游序列、PRD I区,PRD II区和PRD IV区缺失后,LPS同样能诱导EGFP报告基因的表达。只保留PRD区及IFN-β启动子核心区的载体转染Raw264.7细胞后,可以在LPS刺激下诱导EGFP报告基因的高表达。结论构建了能检测IRF-3活性的报告基因载体,为进一步研究LPS激活IRF-3的信号转导通路奠定了基础。 Objective To analyze the regulatory elements that regulates the transcription of murine IFN-β gene and to construct IRF-3 reporter gene. Methods The promoter region of murine IFN-β gene was from murine genomic DNA by PCR and was cloned into promoter-free plasmid pGF3basic/enhancer, amplified upstream of EGFP reporter gene. Vectors with serial deletions at 5＇ side of promoter region were also constructed. Following transient transfection of murine Raw264.7 cells with the vectors and LPS stimulation, the expression of EGFP reporter gene was observed under fluorescent microscopy. Results This study showed that LPS could induce expression of EGFP reporter gene when DNA fragment upstream of PRD region, as well as PRD Ⅰ , Ⅱ and Ⅳ region, were deleted. Conclusion An IRF-3 EGFP reporter gene was established by including only PRD Ⅲ sequence and core region of IFN-β promoter. This reporter gene is an important tool for investigation of LPS-induced IRF-3 signaling transduction pathways.
作者 蒋静 胡福泉 蹇锐 张伟 邓少丽 程小星
机构地区 第三军医大学基础医学部微生物学教研室重庆市微生物工程实验室
出处 《第三军医大学学报》 CAS CSCD 北大核心 2006年第9期896-899,共4页 Journal of Third Military Medical University
基金 国家自然科学基金资助项目(30470094)~~
关键词 IRF-3 IFN-Β 启动子 调控元件 报告基因 IRF-3 IFN-β promoter regulatory element reporter gene
分类号 Q74 [生物学—分子生物学] Q754 [生物学—分子生物学]
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参考文献13

  • 1SAKAGUCHI S,NEGISHI H,ASAGIRI M,et al.Essential role of IRF-3 in lipopolysaccharide-induced interferon-beta gene expression and endotoxin shock[J].Biochem Biophys Res Commun,2003,306 (4):860 -866.
  • 2SERVANT M J,GRANDVAUX N,HISCOTT J.Multiple signaling pathways leading to the activation of interferon regulatory factor 3[J].Biochem Pharmacol,2002,64 (5 -6):985 -992.
  • 3MIYAKE K.Innate recognition of lipopolysaccharide by CD14 and tolllike receptor 4-MD-2:unique roles for MD-2[J].Int Immunopharmacol,2003,3(1):119-128.
  • 4FITZGERALD K A,ROWE D C,BARNES B J,et al.LPS-TLR4 signaling to IRF-3/7 and NF-kappaB involves the toll adapters TRAM and TRIF[J].J Exp Med,2003,198(7):1043 -1055.
  • 5YAMAMOTO M,SATO S,HEMMI H,et al.Role of adaptor TRIF in the MyD88-independent toll-like receptor signaling pathway[J].Science,2003,301(5633):640-643.
  • 6GOODBOURN A,BURSTEIN H,MANIATIS H.The human beta-interferon gene enhancer is under negative control[J].Cell,1986,45 (4):601 -610.
  • 7LEBLANC J F,COHEN L,RODRIGUES M,et al.Synergism between distinct enhanson domains in viral induction of the human beta interferon gene[J].Mol Cell Biol,1990,10(8):3987 -3993.
  • 8DU W,MANIATIS T.An ATF/CREB binding site is required for virus induction of the human interferon beta gene[J].Proc Natl Acad Sci U S A,1992,89(6):2150 -2154.
  • 9TANIGUCHI T,OGASAWARA K,TAKAOKA A,et al,IRF family of transcription factors as regulators of host defense[J].Annu Rev Immunol,2001,(19):623 -655.
  • 10FAN C M,MANIATIS T.Two different virus-inducible elements are required for human beta-interferon gene regulation[J].EMBO J,1989,8 (1):101 -110.

同被引文献25

  • 1张驰宇,徐顺高,黄新祥.一种新颖简便的荧光实时RT-PCR相对定量方法的建立[J].生物化学与生物物理进展,2005,32(9):883-888. 被引量：129
  • 2Moore T C, Cody L, Kumm P M, et al. IRF3 helps control acute TMEV infection through IL-6 expression but contrib- utes to acute hippocampus damage following TMEV infec- tion[J]. Virus Res, 2013, 178(2): 226-233.
  • 3Lin R T, Heylbroeck C, Pitha P M, et al. Virus-dependent phosphorylation of the IRF-3 transcription factor regulates nuclear translocation, transactivation potential, and protea- some-mediated degradation[J]. Mol Cell Biol, 1998, 18(5): 2986-2996.
  • 4Di Paolo N C, Doronin K, Baldwin L K, et al. The transcrip- tion factor IRF3 triggers "defensive suicide" necrosis in re- sponse to viral and bacterial pathogens[J]. Cell Reports, 2013,3(6): 1840-1846.
  • 5Gu M D, Zhang T, Lin W L, et al. Protein phosphatase PP1 negatively regulates the Toll-like receptor- and RIG-I-like receptor-triggered production of type I interferon by inhibit- ing IRF3 phosphorylation at serines 396 and 385 in macro- phage[J]. Cell Signal, 2014, 26(12): 2930-2939.
  • 6Lin L, Pan S, Zhao J Q, et al. HSPD1 interacts with IRF3 to facilitate interferon-beta induction[J]. PloS One, 2014, 9(12): e1 14874.
  • 7Holland J W, Bird S, Williamson B, et al. Molecular charac- terization of IRF3 and IRF7 in rainbow trout, Oncorhynchus mykiss: Functional analysis and transcriptional modulation[J] Mol Immunol, 2008, 46(2): 269-285.
  • 8Song H D, Sun X J, Deng M, et al. Hematopoietic gene ex- pression profile in zebrafish kidney marrow[J]. Proc Natl Acad Sci USA, 2004, 101(46): 16240 -16245.
  • 9Richardson M P, Tay B H, Goh B Y, et al. Molecular cloning and genomic structure of a gene encoding interferon regula- tory factor in the pufferfish (Fugu rubripes)[J]. Mar Bio- technol, 2001, 3(2): 145 -151.
  • 10Sun F, Zhang Y B, Liu T K, et al. Characterization of fish IRF3 as an IFN-inducible protein reveals evolving regulation of IFN response in vertebrates[J]. J Irnmunol, 2010, 185(12): 7573-7582.

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第三军医大学学报
2006年 第9期

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