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Table 2 Requirements of expression plasmids and run-off RNA transcripts for AHSV recovery

From: Requirements and comparative analysis of reverse genetics for bluetongue virus (BTV) and African horse sickness virus (AHSV)

Expression plasmids with authentic ORFs RNA Virus
VP1 VP3 VP4 VP6 VP7 NS1 NS2 Transcripts Recovery
- - - - - - - Capped -
+ + + + + + + Capped +
+ + + + - + + Capped -
+ + + + + - + Capped +
Expression plasmids with optimized ORFs RNA Virus
VP1 VP3 VP4 VP6 VP7 NS1 NS2 Transcripts Recovery
+ + + + + + + Capped +
- + + + + + + Capped -
+ - + + + + + Capped -
+ + - + + + + Capped +
+ + + - + + + Capped +
+ + + + - + + Capped +
+ + + + + - + Capped +
+ + + + + + - Capped -
+ + - - - - + Capped +
+ + + + + + + Uncapped +
+ + - + + + + Uncapped -
+ + + - + + + Uncapped -
+ + + + - + + Uncapped -
+ + + + + - + Uncapped +
+ + - - - - + Uncapped -
+ + + + + + + minSeg-5 -
+ + + + + + + delSeg-5 -
  1. AHSV expression plasmids in the first transfection mix are indicated. In the upper panel, non-optimized ORFs in expression plasmids were used, except for ORF-VP3. In the lower panels, expression plasmids with optimized AHSV-ORFs were used. Then, monolayers were transfected with 10 full length in vitro synthesized capped or uncapped run-off RNA transcripts, except for minSeg-5 RNA (without) and delSeg-5 RNA (Seg-5 RNA with a small out-of-frame deletion). Virus recovery was microscopically determined by CPE and confirmed by IPMA, and indicated as positive (+) or negative (-)