通过 Tn7 介导的位点特异性转座将外来基因插入猿水痘病毒基因组

IF 2.2 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS Journal of virological methods Pub Date : 2024-04-06 DOI:10.1016/j.jviromet.2024.114936

Wayne L. Gray

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引用次数: 0

摘要

利用 Tn7 转座方法将外来基因定点插入猿水痘病毒（SVV）基因组，SVV 是非人灵长类动物猿水痘的病原体。将严重急性呼吸系统综合征冠状病毒（SARS-CoV-2）的核头状（N）基因和穗状基因的受体结合域（RBD）插入克隆到细菌人工染色体的 SVV 基因组 ORF 14 区域，然后转染到 Vero 细胞中，产生具有传染性的重组 SVV（rSVV）。rSVV在感染的Vero细胞中高效复制，并通过免疫印迹和免疫荧光检测表达N和RBD抗原。Tn7 介导的转座为构建可用作减毒活疫苗的 rSVV 提供了一种快速有效的方法。

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Insertion of foreign genes into the simian varicella virus genome by Tn7-mediated site-specific transposition

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.

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来源期刊

Journal of virological methods 医学-病毒学

CiteScore

5.80

自引率

0.00%

发文量

209

审稿时长

41 days

期刊介绍： The Journal of Virological Methods focuses on original, high quality research papers that describe novel and comprehensively tested methods which enhance human, animal, plant, bacterial or environmental virology and prions research and discovery. The methods may include, but not limited to, the study of: Viral components and morphology- Virus isolation, propagation and development of viral vectors- Viral pathogenesis, oncogenesis, vaccines and antivirals- Virus replication, host-pathogen interactions and responses- Virus transmission, prevention, control and treatment- Viral metagenomics and virome- Virus ecology, adaption and evolution- Applied virology such as nanotechnology- Viral diagnosis with novelty and comprehensive evaluation. We seek articles, systematic reviews, meta-analyses and laboratory protocols that include comprehensive technical details with statistical confirmations that provide validations against current best practice, international standards or quality assurance programs and which advance knowledge in virology leading to improved medical, veterinary or agricultural practices and management.