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融合铁蛋白的猪瘟病毒囊膜蛋白E2的表达及鉴定 被引量:1
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作者 魏珍珍 曾振 +5 位作者 杜梦潭 刘兴健 张志芳 胡小元 李轶女 易咏竹 《生物技术进展》 2020年第4期386-392,共7页
目前,传统的灭活猪瘟疫苗及弱毒(减毒)猪瘟疫苗存在免疫保护期短、毒力返强等缺点,而蛋白源性亚单位疫苗可以很好的解决上述问题。铁蛋白24个亚基可以自组装成稳定的多聚体纳米颗粒,成为理想的抗原呈递载体和疫苗开发平台。基于此,将铁... 目前,传统的灭活猪瘟疫苗及弱毒(减毒)猪瘟疫苗存在免疫保护期短、毒力返强等缺点,而蛋白源性亚单位疫苗可以很好的解决上述问题。铁蛋白24个亚基可以自组装成稳定的多聚体纳米颗粒,成为理想的抗原呈递载体和疫苗开发平台。基于此,将铁蛋白与具有较强免疫原性的猪瘟病毒囊膜蛋白E2(具有高保守性的保护性抗原)结合,构建融合基因E2-Fe,将该基因克隆到pET28a原核表达载体中,转化至大肠杆菌(Escherichia coli)BL21(DE3)感受态细胞中进行诱导条件的探索及其高效表达,再将表达的融合蛋白进行镍柱纯化、质谱分析、Western-blotting验证及电镜检测。结果显示,融合基因E2-Fe已成功克隆到pET-28a载体上,0.50%乳糖诱导6 h为融合蛋白E2-Fe表达的最优条件;质谱分析与Western-blotting均显示融合蛋白的大小约为51 kD,与理论值相符;电镜观察到的纳米颗粒直径约为20 nm。结果表明,融合铁蛋白的猪瘟病毒囊膜蛋白E2在大肠杆菌中得到高效表达,经镍柱纯化后可获得数量可观的自组装纳米颗粒抗原,为研究新的猪瘟疫苗拓宽了研究思路。 展开更多
关键词 猪瘟病毒囊膜蛋白E2 纳米颗粒抗原 铁蛋白
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Preparation of Cu nanoparticles with ascorbic acid by aqueous solution reduction method 被引量:7
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作者 刘清明 Takehiro YASUNAMI +1 位作者 Kensuke KURUDA Masazumi OKIDO 《Transactions of Nonferrous Metals Society of China》 SCIE EI CAS CSCD 2012年第9期2198-2203,共6页
Cu nanoparticles were prepared by reducing Cu2+ ions with ascorbic acid through aqueous solution reduction method. The effects of solution pH and average size of Cu2O particles on the preparation of Cu nanoparticles ... Cu nanoparticles were prepared by reducing Cu2+ ions with ascorbic acid through aqueous solution reduction method. The effects of solution pH and average size of Cu2O particles on the preparation of Cu nanoparticles were investigated. Cu particles were prepared at pH 3, 5 or 7, with the smallest Cu particles obtained at pH 7. However, Cu particles could not be prepared at pH 9 or 11. The average size of Cu2O particles can affect that of Cu particles. Larger Cu2O particles result in larger Cu particles. In addition, experiments were conducted to explore the reaction process by measuring the X-ray diffraction (XRD) patterns of specimens collected at different time points during the reaction. It was found that Cu(OH)2 was initially formed as a precursor, followed by the formation of Cu2O, which was finally reduced to Cu particles. 展开更多
关键词 Cu nanoparticles aqueous solution reduction method ascorbic acid intermediate product
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