报道了短尾蝮蛇Gloydius brevicaudus干扰素调节因子2基因(IRF-2)的克隆和cDNA全序列测定分析。目前除在爬行类外,IRF-2基因在大部分的脊椎动物如鱼类、两栖类、鸟类和哺乳类都有报道。为了获得爬行类IRF-2基因的全序列,从短尾蝮蛇的肾...报道了短尾蝮蛇Gloydius brevicaudus干扰素调节因子2基因(IRF-2)的克隆和cDNA全序列测定分析。目前除在爬行类外,IRF-2基因在大部分的脊椎动物如鱼类、两栖类、鸟类和哺乳类都有报道。为了获得爬行类IRF-2基因的全序列,从短尾蝮蛇的肾、脾、肝和肠4种组织中使用Trizol试剂盒提取了总RNA。从已知的IRF-2基因序列比对设计了简并引物来扩增保守片段。最后使用RACE方法得到IRF-2的cDNA全序列。结果显示短尾蝮蛇的IRF-2基因开放阅读框包含有978个核苷酸,编码326个氨基酸,其3′UTR包含137个核苷酸。与脊椎动物其他四足动物序列比对分析还发现,短尾蝮蛇的IRF-2基因和推导的氨基酸序列都非常保守,与大部分四足动物的IRF-2相似。从其氨基酸序列中可以辨别出DBD、IDA2和transactivating dom ain等大部分元件和阻遏模体。展开更多
Interferon Regulatory Factor-2 (IRF-2) belongs to IRF family, was identified as a mammalian transcription factor involved in Interferon beta (IFNβ) gene regulation. Besides that IRF-2 is involved in immunomodulation,...Interferon Regulatory Factor-2 (IRF-2) belongs to IRF family, was identified as a mammalian transcription factor involved in Interferon beta (IFNβ) gene regulation. Besides that IRF-2 is involved in immunomodulation, hematopoietic differentiation, cell cycle regulation and oncogenesis. We have done molecular sub-cloning and expression of recombinant murine IRF-2 as GST (Glutathione-S-Transferase)- IRF-2 fusion protein in E. coli/XL-1blue cells. Recombinant IRF-2 with GST moiety at N-terminus expressed as GST-IRF-2 (~66 kd) in E. coli along with different low molecular mass degradation products revealed approximately 30, 42, 60 and 62 kd by SDS-PAGE and Western blot, respectively. We further confirm that degradation takes place at C-terminus of the fusion protein not at N-terminus as anti-GST antibody was detecting all bands in the immunoblot. The recombinant IRF-2 was biologically active along with their degradation products in terms of their DNA binding activity as assessed by Electrophoretically Mobility Shift Assay (EMSA). We observed three different molecular mass DNA/protein complexes (1 - 3) with Virus Response Element (VRE) derived from human Interferon IFNβ gene and five different molecular mass complexes (1 - 5) with IRF-E motif (GAAAGT)4 in EMSA gel. GST only expressed from empty vector did not bind to these DNA elements. To confirm that the binding is specific, all complexes were competed out completely when challenged with 100-X fold molar excess of IRF-E oligo under cold competition. It means degradation products along with full-length protein are able to interact with VREβ as well as IRF-E motif. This means degradation products may regulate the target gene (s) activation/repression via interacting with VRE/IRF-E.展开更多
针对装配式钢筋模板一体化(Integration of reinforcement and framework,IRF)体系采用硫铝酸盐水泥存在成本高、性能调控难度大等问题,基于最紧密堆积理论设计了满足装配式IRF体系性能要求的P·Ⅱ水泥砂浆配合比,研究了该砂浆在不...针对装配式钢筋模板一体化(Integration of reinforcement and framework,IRF)体系采用硫铝酸盐水泥存在成本高、性能调控难度大等问题,基于最紧密堆积理论设计了满足装配式IRF体系性能要求的P·Ⅱ水泥砂浆配合比,研究了该砂浆在不同CO_(2)养护温度(40℃、60℃)和养护压强(0.3、0.5、0.8 MPa)下的力学性能、水化热、水化产物和孔结构的变化规律,并与60℃、常压蒸汽养护组进行了对比。结果表明:60℃下,与蒸汽养护相比,CO_(2)养护有效促进了砂浆早期抗压强度发展,建议CO_(2)养护压强为0.5 MPa;CO_(2)养护延长了胶凝材料的水化诱导期,降低了水化放热速率峰值和24 h水化放热量;养护压强对胶凝材料水化动力学的影响不显著;用适量辅助胶凝材料替代水泥细化了硬化水泥浆体的孔结构,且碳减排效果显著。展开更多
文摘报道了短尾蝮蛇Gloydius brevicaudus干扰素调节因子2基因(IRF-2)的克隆和cDNA全序列测定分析。目前除在爬行类外,IRF-2基因在大部分的脊椎动物如鱼类、两栖类、鸟类和哺乳类都有报道。为了获得爬行类IRF-2基因的全序列,从短尾蝮蛇的肾、脾、肝和肠4种组织中使用Trizol试剂盒提取了总RNA。从已知的IRF-2基因序列比对设计了简并引物来扩增保守片段。最后使用RACE方法得到IRF-2的cDNA全序列。结果显示短尾蝮蛇的IRF-2基因开放阅读框包含有978个核苷酸,编码326个氨基酸,其3′UTR包含137个核苷酸。与脊椎动物其他四足动物序列比对分析还发现,短尾蝮蛇的IRF-2基因和推导的氨基酸序列都非常保守,与大部分四足动物的IRF-2相似。从其氨基酸序列中可以辨别出DBD、IDA2和transactivating dom ain等大部分元件和阻遏模体。
文摘Interferon Regulatory Factor-2 (IRF-2) belongs to IRF family, was identified as a mammalian transcription factor involved in Interferon beta (IFNβ) gene regulation. Besides that IRF-2 is involved in immunomodulation, hematopoietic differentiation, cell cycle regulation and oncogenesis. We have done molecular sub-cloning and expression of recombinant murine IRF-2 as GST (Glutathione-S-Transferase)- IRF-2 fusion protein in E. coli/XL-1blue cells. Recombinant IRF-2 with GST moiety at N-terminus expressed as GST-IRF-2 (~66 kd) in E. coli along with different low molecular mass degradation products revealed approximately 30, 42, 60 and 62 kd by SDS-PAGE and Western blot, respectively. We further confirm that degradation takes place at C-terminus of the fusion protein not at N-terminus as anti-GST antibody was detecting all bands in the immunoblot. The recombinant IRF-2 was biologically active along with their degradation products in terms of their DNA binding activity as assessed by Electrophoretically Mobility Shift Assay (EMSA). We observed three different molecular mass DNA/protein complexes (1 - 3) with Virus Response Element (VRE) derived from human Interferon IFNβ gene and five different molecular mass complexes (1 - 5) with IRF-E motif (GAAAGT)4 in EMSA gel. GST only expressed from empty vector did not bind to these DNA elements. To confirm that the binding is specific, all complexes were competed out completely when challenged with 100-X fold molar excess of IRF-E oligo under cold competition. It means degradation products along with full-length protein are able to interact with VREβ as well as IRF-E motif. This means degradation products may regulate the target gene (s) activation/repression via interacting with VRE/IRF-E.
文摘针对装配式钢筋模板一体化(Integration of reinforcement and framework,IRF)体系采用硫铝酸盐水泥存在成本高、性能调控难度大等问题,基于最紧密堆积理论设计了满足装配式IRF体系性能要求的P·Ⅱ水泥砂浆配合比,研究了该砂浆在不同CO_(2)养护温度(40℃、60℃)和养护压强(0.3、0.5、0.8 MPa)下的力学性能、水化热、水化产物和孔结构的变化规律,并与60℃、常压蒸汽养护组进行了对比。结果表明:60℃下,与蒸汽养护相比,CO_(2)养护有效促进了砂浆早期抗压强度发展,建议CO_(2)养护压强为0.5 MPa;CO_(2)养护延长了胶凝材料的水化诱导期,降低了水化放热速率峰值和24 h水化放热量;养护压强对胶凝材料水化动力学的影响不显著;用适量辅助胶凝材料替代水泥细化了硬化水泥浆体的孔结构,且碳减排效果显著。
