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目的构建多房棘球绦虫(Em)重组双歧杆菌(Bb)-Em14-3-3疫苗,并研究Em14-3-3分子在大肠埃希菌BL21(DE3)中的表达效率。方法通过PCR扩增Em14-3-3抗原编码基因;将该基因定向克隆于含有谷胱甘肽-S-转移酶(GST)基因的大肠埃希菌-双歧杆菌穿梭表达载体pGEX-1λT,构建重组质粒pGEX-Em14-3-3;用电穿孔法将该质粒导入Bb,构建多房棘球绦虫重组Bb-Em14-3-3疫苗。经PCR和酶切鉴定后以IPTG诱导表达Em14-3-3/GST融合蛋白;SDS-PAGE及Western blot对表达产物进行鉴定。结果PCR成功扩增出530 bp的Em14-3-3基因;双酶切证实Em14-3-3基因插入pGEX-1λT中;PCR证实重组Bb-Em14-3-3疫苗构建成功;SDS-PAGE及Western blot分析显示重组质粒转化宿主菌在IPTG诱导下高效表达了Em14-3-3/GST融合蛋白,表达效率为23%。结论成功构建了多房棘球绦虫重组Bb-Em14-3-3疫苗,重组质粒pGEX-Em14-3-3在大肠杆菌中获得了高效表达,Western blot结果提示表达出的Em14-3-3重组蛋白具有特异抗原性。 相似文献
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本文在充分分析我国西部地区综合运输服务发展现状及存在问题的基础上,分析“十四五”期间发展形势与趋势,剖析西部地区与东部地区之间存在的差距,提出了打造适应需求的基础设施体系、构建便捷品质的客运服务体系、构建集约高效的货运服务体系以及提升科技创新活力推动运输服务升级等四项举措,为西部地区综合运输服务水平提升和高质量发展提供参考。 相似文献
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14MnNbq钢板厚度方向性能试验研究 总被引:3,自引:0,他引:3
介绍对天兴洲大桥钢梁用14MnNbq钢进行的板厚方向拉伸试验,以及统计分析,研究结果表明,当含硫量为0.01%时,预估断面收缩率ψZ≈43%,大于Z35级标准。因此,天兴洲大桥钢板订货时可不必另提补充条件,使用时,需对所用处的钢板进行ψZ试验并进行探伤,满足Z35级的要求。 相似文献
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应用3H-TdR及14C-UR参入淋巴细胞的双标记方法,在体外培养条件下,研究多抗甲素对人淋巴细胞DNA及RNA合成功能的影响。结果表明,在多抗甲素的浓度为0.1~10mg/L时,可显著增强淋巴细胞DNA及RNA的合成功能;当多抗甲素的浓度增至50~100mg/L时,对淋巴细胞合成DNA及RNA的能力呈现抑制作用,3H及14C标记物的参入计数减少。多抗甲素对淋巴细胞DNA及RNA合成功能的影响呈现双相作用。 相似文献
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Hui Jin Hai-Tao Hu Wei-Xi Wang Gai-Feng Feng Zhao-Hui Liu Wei-Na Yang 《西安交通大学学报(英文版)》2010,22(2):105-110
Objective To investigate the expression of neuroprotective peptide [Gly14]-Humanin (HNG) in eukaryotic cells by gene engineering technique and analyze its biological activity. Methods By means of asymmetrical primer/template, double stranded cDNA of HNG with FLAG in its C-terminal was obtained, which was cloned into the plasmid pcDNA3.1(-), and the resultant recombinant vector pcDNA3.1(-)/HNG-FLAG was transfected into PC12 cells. At the same time, the recombinant vector pcDNA3.1(-)/EGFP was transfected to control the efficiency of transfection. The expression of HNG in the cells was determined by immunocytochemistry. In order to analyze the biological activity of the expressed HNG, 25μM Aβ25-35 peptide was added to the culture medium of the transfected cells for 24h, then cell morphology, MTT assay and Hoechst 33258 staining were observed. Results The eukaryotic expression vector of pcDNA3.1(-)/HNG-FLAG was identified by enzyme digestion and sequencing. HNG was highly expressed in PC12 cells. After exposure of PC12 cells to 25μM Aβ25-35 for 24h, cell viability decreased to (65.8±5.3)%, and the dystrophic changes of neuritis and nuclei condensation were obvious. When cells were pre-transfected with pcDNA3.1(-)/HNG-FLAG, Aβ25-35-induced cell death and morphological changes of cells and nuclei were suppressed. In contrast, pre-transfected with empty vector did not protect cells from Aβ25-35-induced toxicity. Conclusion The eukaryotic expression vector for FLAG-tagged HNG was successfully constructed and expressed in PC12 cells. Expressed HNG has biological activity. 相似文献
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14MnNbq钢要求屈服强度大于或等于390MPa,同时要满足低温抗断要求的可焊性和韧性。为了满足上述性能,提出了其化学成分设计原则,并进一步提出14MnNbq钢性能优化的技术条件,即化学成分优化和力学性能优化。14MnNbq钢要广泛应用于桥梁建设,尚需按照结构设计形式、结构制造工艺、荷载作用特点和使用环境进行大量的实验研究,以满足结构安全使用和结构使用寿命的要求。 相似文献
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