PDF(Pubmed)
Abstract：
A Tn7-transposition approach was utilized for site-specific insertion of foreign genes into the genome of simian varicella virus (SVV), the causative agent of simian varicella in nonhuman primates. The severe acute respiratory syndrome coronavirus (SARS-CoV-2) nucleocapsid (N) gene and receptor binding domain (RBD) of the spike gene were inserted into the ORF 14 region of the SVV genome cloned into a bacterial artificial chromosome and then transfected into Vero cells to generate the infectious recombinant SVV (rSVV). The rSVV replicated efficiently in infected Vero cells and expressed the N and RBD antigens as indicated by immunoblot and immunofluorescence assays. Tn7-mediated transposition provides a rapid and efficient method for constructing rSVVs which may be evaluated as live-attenuated vaccines.
摘要：
Tn7转座方法用于将外源基因位点特异性插入猿猴水痘病毒（SVV）的基因组中，非人灵长类动物中猿猴水痘的病原体。将严重急性呼吸综合征冠状病毒（SARS-CoV-2）核衣壳（N）基因和刺突基因的受体结合域（RBD）插入克隆到细菌人工染色体中的SVV基因组的ORF14区域，然后转染到Vero细胞中，以产生感染性重组SVV（rSVV）。如免疫印迹和免疫荧光测定所示,rSVV在感染的Vero细胞中有效复制并表达N和RBD抗原。Tn7介导的转座为构建rSVV提供了一种快速有效的方法，可将其评估为减毒活疫苗。
