Rescue of viral haemorrhagic septicaemia virus minigenomes by helper virus.
A mammalian expression vector containing the bacterial chloramphenicol acetyltransferase (CAT) gene was used to demonstrate that CAT could be successfully used as a reporter system in fish cells growing at low temperatures. We then constructed a viral haemorrhagic septicaemia virus (VHSV) minigenome by cloning the CAT reporter gene between the viral leader and trailer sequences. This construct was used in transfection experiments with helper VHSV to demonstrate that the minigenome can be encapsidated and transcribed by helper virus proteins. In addition, passaging of viruses collected from cells expressing the minigenome showed that the minigenome was being packaged and replicated in the presence of helper virus. These experiments provide the initiating steps for a reverse genetics system for VHSV.
Betts
A M
AM
The Centre for the Environment, Fisheries and Aquaculture Science, Weymouth Laboratory, Barrack Road, Weymouth, DT4 8UB, Dorset, UK. a.m.betts@cefas.co.uk
Stone
D M
DM
D000818:Animals; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D005393:Fish Diseases / Q000821:virology; D005399:Fishes; D017930:Genes, Reporter; D016679:Genome, Viral; D006378:Helper Viruses / Q000235:genetics* / Q000502:physiology; D012209:Rhabdoviridae / Q000235:genetics* / Q000502:physiology*; D018353:Rhabdoviridae Infections / Q000662:veterinary / Q000821:virology; D014158:Transcription, Genetic; D014162:Transfection; D014779:Virus Replication
D015500:Chloramphenicol O-Acetyltransferase