A Novel Strategy of US3 Codon De-Optimization for Construction of an Attenuated Pseudorabies Virus against High Virulent Chinese Pseudorabies Virus Variant.

ABSTRACT: In this study, we applied bacterial artificial chromosome (BAC) technology with PRV^ΔTK/gE/gI as the base material to replace the first, central, and terminal segments of the US3 gene with codon-deoptimized fragments via two-step Red-mediated recombination in E. coli GS1783 cells. The three constructed BACs were co-transfected with gI and part of gE fragments carrying homologous sequences (gI+gE'), respectively, in swine testicular cells. These three recombinant viruses with US3 codon de-optimization ((PRV^{ΔTK&gE-US3deop-1}, PRV^{ΔTK&gE-US3deop-2}, and PRV^{ΔTK&gE-US3deop-3}) were obtained and purified. These three recombinant viruses exhibited similar growth kinetics to the parental AH02LA strain, stably retained the deletion of TK and gE gene fra

SUBMITTER: Xu M 

PROVIDER: S-EPMC10458909 | biostudies-literature | 2023 Jul

REPOSITORIES: biostudies-literature