在昆虫细胞中生产严重急性呼吸综合征冠状病毒2（SARS-CoV-2）病毒样颗粒
Production of SARS-CoV-2 Virus-Like Particles in Insect Cells.
作者信息
Mi Youjun, Xie Tao, Zhu Bingdong, Tan Jiying, Li Xuefeng, Luo Yanping, Li Fei, Niu Hongxia, Han Jiangyuan, Lv Wei, Wang Juan
机构信息
Lanzhou Center for Tuberculosis Research and Gansu Provincial Key Laboratory of Evidence Based Medicine and Clinical Translation, School of Basic Medical Sciences, Lanzhou University, Lanzhou 730070, China.
Institute of Immunology, School of Basic Medicine, Lanzhou University, Lanzhou 730070, China.
出版信息
Vaccines (Basel). 2021 May 26;9(6):554. doi: 10.3390/vaccines9060554.
Coronavirus disease (COVID-19) causes a serious threat to human health. Virus-like particles (VLPs) constitute a promising platform in SARS-CoV-2 vaccine development. In this study, the E, M, and S genes were cloned into multiple cloning sites of a new triple expression plasmid with one p10 promoter, two pPH promoters, and three multiple cloning sites. The plasmid was transformed into DH10 Bac competent cells to obtain recombinant bacmid. Then the recombinant bacmid was transfected in ExpiSf9 insect cells to generate recombinant baculovirus. After ExpiSf9 cells infection with the recombinant baculovirus, the E, M, and S proteins were expressed in insect cells. Finally, SARS-CoV-2 VLPs were self-assembled in insect cells after infection. The morphology and the size of SARS-CoV-2 VLPs are similar to the native virions.
冠状病毒病（COVID-19）对人类健康构成严重威胁。病毒样颗粒（VLPs）是严重急性呼吸综合征冠状病毒2（SARS-CoV-2）疫苗研发中一个很有前景的平台。在本研究中，E、M和S基因被克隆到一个新的三表达质粒的多个克隆位点中，该质粒具有一个p10启动子、两个pPH启动子和三个多个克隆位点。将该质粒转化到DH10 Bac感受态细胞中以获得重组杆粒。然后将重组杆粒转染到ExpiSf9昆虫细胞中以产生重组杆状病毒。在用重组杆状病毒感染ExpiSf9细胞后，E、M和S蛋白在昆虫细胞中表达。最后，感染后SARS-CoV-2 VLPs在昆虫细胞中自组装。SARS-CoV-2 VLPs的形态和大小与天然病毒粒子相似。
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