Complementation studies were carried out, using temperature-sensitive (t-s) mutants of bluetongue virus (BTV). The results proved to be inconclusive as only low indices of complementation were obtained. No discrepancy was found between the previous classification of these mutants in 6 recombination classes and the complementation data recorded. In general, the t-s mutants require a latent growth period of 16-20 h at 28 °C and maximum titres can be demonstrated 40-48 h post-infection. One mutant, (F211), however, consistently had a growth lag phase of 32 h. Mutants of the 6 recombination groups were further classified into 2 groups by temperature-shift studies. One class of mutants expressed their t-s lesion prior to 24 h and the other class only after 24 h post-infection. Mutant F73 was found to be defective in its ability to synthesize ssRNA at a late stage in the replication cycle at the non-permissive temperature.
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