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HIV-1 CAP2NC蛋白的表达及体外自组装
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作者 白诗梦 张芝晴 +7 位作者 乔佳明 沈鸿霖 黄芳 高双全 李少勇 李少伟 夏宁邵 顾颖 《生物工程学报》 CAS CSCD 北大核心 2018年第4期586-593,共8页
构建并表达HIV-1 CAP2NC蛋白,探索其体外自组装条件。通过PCR技术扩增HIV-1(NL4-3毒株)CAP2NC基因片段,并将其连接到原核表达载体pTO-T7,获得重组质粒pTO-T7-CAP2NC,然后转化至大肠杆菌BL21(DE3)菌株,经疏水层析纯化后获得重组蛋白CAP2N... 构建并表达HIV-1 CAP2NC蛋白,探索其体外自组装条件。通过PCR技术扩增HIV-1(NL4-3毒株)CAP2NC基因片段,并将其连接到原核表达载体pTO-T7,获得重组质粒pTO-T7-CAP2NC,然后转化至大肠杆菌BL21(DE3)菌株,经疏水层析纯化后获得重组蛋白CAP2NC。SDS-PAGE结果表明,重组蛋白CAP2NC可在大肠杆菌可溶高效表达,经纯化后纯度约为95%。ELISA检测表明重组蛋白CAP2NC可被HIV-1衣壳蛋白特异性单克隆抗体识别,具有较好反应活性。重组蛋白透析后在非原性SDS-PAGE中呈现为多种聚体形式。分子筛排阻层析分析CAP2NC蛋白透析后可进行组装,负染电镜进一步观察显示CAP2NC蛋白在RNA存在条件下,可形成空心管状颗粒,其形态结构与HIV-1病毒衣壳体外自组装形成的类似。上述结果表明HIV-1 CAP2NC蛋白具有体外自组装的性质,为进一步在体外研究非成熟病毒样颗粒结构奠定基础。 展开更多
关键词 HIV-1 CAP2NC蛋白 自组装
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AN EXTRACTION ALGORITHM FOR A SET OF ELEMENTARY SIPHONS BASED ON MIXED-INTEGER PROGRAMMING 被引量:1
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作者 shaoyong li Zhiwu li +2 位作者 Hesuan HU Abdulrahman AI-AHMARI Aimin AN 《Journal of Systems Science and Systems Engineering》 SCIE EI CSCD 2012年第1期106-125,共20页
Elementary siphons are useful in the development of a deadlock prevention policy for a discrete event system modeled with Petri nets. This paper proposes an algorithm to iteratively extract a set of elementary siphons... Elementary siphons are useful in the development of a deadlock prevention policy for a discrete event system modeled with Petri nets. This paper proposes an algorithm to iteratively extract a set of elementary siphons in a class of Petri nets, called system of simple sequential processes with resources (S3pR). At each iteration, by a mixed-integer programming (MIP) method, the proposed algorithm finds a maximal unmarked siphon, classifies the places in it, extracts an elementary siphon from the classified places, and adds a new constraint in order to extract the next elementary siphon. This algorithm iteratively executes until no new unmarked siphons can be found. It finally obtains a unique set of elementary siphons and avoids a complete siphon enumeration. A theoretical analysis and examples are given to demonstrate its efficiency and practical potentials. 展开更多
关键词 Petri net flexible manufacturing system deadlock prevention mixed integer programming elementary siphon
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