Development of EGFP/GLUC-tagged Sindbis-like virus XJ-160.

Based on an infectious clone of Sindbis-like virus XJ-160, recombinant vectors containing a reporter gene (enhanced green fluorescence protein [EGFP] or Gaussia luciferase [GLUC]) were constructed by placing the reporter gene cassette containing the subgenomic promoter behind the 3' terminus of the viral structural protein gene. EGFP/GLUC-tagged Sindbis-like viruses were rescued in BHK-21 cells transfected with transcripts produced from the recombinant vectors. EGFP expression and strong luciferase activity were detected in BHK-21 cells infected with repeated passages of the EGFP/GLUC-tagged viruses, revealing the genetic stability of the chimeric viruses. The EGFP/GLUC-tagged Sindbis viruses reported will contribute to the assessment of viral replication and proliferation, tracking and elucidating Alphavirus-host interactions, and screening for antiviral compounds.