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Abstract：
This study was conducted to efficiently produce virus-like particles (VLPs) of enterovirus 71 (EV71), a causative virus of hand, foot, and mouth disease (HFMD). The expression level of the P1 precursor, a structural protein of EV71, was modified to increase VLP production, and the optimal expression level and duration of the 3CD protein for P1 cleavage were determined. The expression level and duration of 3CD were controlled by the p10 promoter, which was weakened by repeated burst sequence (BS) applications, as well as the OpIE2 promoter, which was weakened by the insertion of random untranslated region sequences of various lengths. The cleavage and production efficiency of the P1 precursor were compared based on the expression time and level of 3CD, revealing that the p10-BS5 promoter with four repeated BSs was the most effective. When P1 and 3CD were expressed using the hyperexpression vector and the p10-BS5 promoter, high levels of structural protein production and normal HFMD-VLP formation were observed, respectively. This study suggests that the production efficiency of HFMD-VLPs can be significantly enhanced by increasing the expression of the P1 precursor and controlling the amount and duration of 3CD expression.
摘要：
本研究旨在高效生产肠道病毒71型(EV71)的病毒样颗粒(VLP),手的致病病毒，脚,和口蹄疫（HFMD）。P1前体的表达水平，EV71的结构蛋白被修饰以增加VLP的产生，并确定了P1裂解的3CD蛋白的最佳表达水平和持续时间。3CD的表达水平和持续时间由p10启动子控制，它被重复的突发序列(BS)应用削弱了，以及OpIE2启动子，通过插入各种长度的随机非翻译区序列而被削弱。根据3CD的表达时间和水平比较P1前体的裂解和生产效率，揭示了具有四个重复BS的p10-BS5启动子是最有效的。当使用过表达载体和p10-BS5启动子表达P1和3CD时，观察到高水平的结构蛋白产生和正常的HFMD-VLP形成，分别。这项研究表明，通过增加P1前体的表达并控制3CD表达的量和持续时间，可以显着提高HFMD-VLP的生产效率。
