The NV genes of fish rhabdoviruses: development of RNase protection assays for rapid assessment of genetic variation.
The genomic RNA of the fish rhabdovirus infectious hematopoietic necrosis virus (IHNV) contains a small NV gene which is not present in any other rhabdoviruses characterized to date. We have constructed a plasmid which carries a full length cDNA copy of the IHNV NV gene between 2 RNA polymerase promoters such that plus-sense and minus-sense RNA transcripts of the IHNV NV gene can be synthesized. These were used to develop an RNase protection assay which was capable of detecting as little as 1-5 ng of NV mRNA or genomic RNA. RNA from tissue culture cells infected with 3 different strains of IHNV showed distinct, diagnostic cleavage patterns reflecting sequence variation in their NV genes.
Kurath
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G
National Biological Service, Northwest Biological Science Center, Seattle, WA 98115, USA.
Higman
K H
KH
Björklund
H V
HV
D000818:Animals; D001345:Autoradiography / Q000379:methods; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods; D016384:Consensus Sequence; D017931:DNA Primers; D005399:Fishes / Q000821:virology*; D005814:Genes, Viral; D014644:Genetic Variation; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D012367:RNA, Viral / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D012209:Rhabdoviridae / Q000235:genetics*; D012260:Ribonucleases; D012680:Sensitivity and Specificity; D014158:Transcription, Genetic; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics*
D017931:DNA Primers; C096112:NV protein, Rhabdovirus; D012333:RNA, Messenger; D012367:RNA, Viral; D014764:Viral Proteins; D012260:Ribonucleases