title: [Rapid selection of recombinant orf virus expression vectors using green fluorescent protein].
abstract: OBJECTIVE
  To construct a universal, highly attenuated orf virus expression vector for exogenous genes using green fluorescent protein (GFP) as the reporter gene.
  
  
  METHODS
  The flanking regions of the ORFV132 of orf virus DNA were amplified by PCR to construct the shuttle plasmid pSPV-132LF-EGFP-132RF. The shuttle plasmid was transfected into OFTu cells and GFP was incorporated into orf virus IA82Delta 121 by homologous recombination. The recombinant IA82Delta121-V was selected by green fluorescent signal. The deletion gene was identified by PCR and sequencing. The effects of ORFV132 knockout were evaluated by virus titration and by observing the proliferation of the infected vascular endothelial cells in vitro.
  
  
  RESULTS
  The recombinant orf virus IA82Delta121-V was obtained successfully and quickly, and the deletion of ORFV132 did not affect the replication of the virus in vitro but reduced its virulence.
  
  
  CONCLUSIONS
  Green fluorescent protein is a selectable marker for rapid, convenient and stable selection of the recombinant viruses. Highly attenuated recombinant orf virus IA82Delta121-V can serve as a new expression vector for exogenous genes.
authors.affiliation: Department of Laboratory Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China.E-mail: 452398956@qq.com.
authors.forename: Jiachun
authors.initials: J
authors.lastname: Zhang
authors.forename: Xianfeng
authors.initials: X
authors.lastname: Guo
authors.forename: Min
authors.initials: M
authors.lastname: Zhang
authors.forename: Feifan
authors.initials: F
authors.lastname: Wu
authors.forename: Yongzheng
authors.initials: Y
authors.lastname: Peng
mesh_terms: D002478:Cells, Cultured; D042783:Endothelial Cells / Q000378:metabolism; D017353:Gene Deletion; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D009923:Orf virus; D010957:Plasmids; D014162:Transfection
chemical_list: D049452:Green Fluorescent Proteins