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Transcription in vitro of Infectious Haematopoietic Necrosis Virus, a Fish Rhabdovirus

Gael Kurath

Department of Microbiology, Oregon State University, Corvallis, Oregon 97331-3804, U.S.A.

Transcription in vitro by the RNA polymerase of infectious haematopoietic necrosis virus (IHNV), a salmonid rhabdovirus, was investigated using different reaction conditions to maximize RNA synthesis. The use of HEPES buffer rather than Tris buffer, and the addition of S-adenosyl-L-methionine to the reactions resulted in a sixfold increase in RNA synthetic activity to 6400 pmol UMP incorporated/mg viral protein/hour. The RNA transcripts produced in this system contained polyadenylated species which co-migrated with IHNV mRNA species 2, 3, 4 and 5 from IHNV-infected cells. The transcripts were shown to be functional mRNA species by their ability to direct the synthesis of viral proteins in vitro.

Keywords: IHNV, transcription in vitro, fish rhabdovirus polymerase

Present address: Department of Plant Pathology, Cornell University, Ithaca, New York 14853, U.S.A.

Received 7 October 1986; accepted 26 February 1987.