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Production of Replication-Defective Retrovirus by Transient Transfection of 293T cells
Published on: December 4, 2007
In vitro transcriptase deficiency of temperature-dependent host range mutants of Chandipura virus
Abstract:
Ten of 12 Chandipura virus tdCE mutants, which exhibit temperature-dependent restriction of growth in chick embryo (CE) cells but not in BS-C-1 cells, showed deficient transcriptase activity in vitro at 39 degrees C relative to wild-type virus. A gradation in transcriptional activity at 39 degrees C in vitro was observed. Reversion of the tdCE phenotype to unrestricted growth in CE cells at 39 degrees C was accompanied by partial restoration of normal transcriptase activity at 39 degrees C, suggesting that reversion was mediated by either extragenic or intragenic suppression. Viral protein synthesis was reduced or absent in CE cells at 39 degrees C indicating that transcription was also defective in vivo under these conditions. Induction of heat-shock proteins in CE cells at 39 degrees C occurred normally in tdCE mutant-infected cells and RNA methylation in vitro was unaffected.

