The complete genome of snakehead rhabdovirus (SHRV) was cloned and
molecularly characterized. This was initially accomplished through the sequence
determination of its glycoprotein gene and the phylogenetic analysis of this gene with
orthologous genes from other rhabdoviruses. The phylogenetic analysis revealed that
SHRV groups with viruses of the genus Novirhabdovirus. The full-length glycoprotein
was expressed in mammalian cells to investigate its potential use in the production of
pseudotyped retroviruses.
The sequence of the entire SHRV genome of 11.6 kb was determined, and all
encoded proteins, intergenic transcriptional control motifs, and the leader and trailer
regions were identified. The genome was found to encode six proteins including a
nucleoprotein, a phosphoprotein, a matrix protein, a glycoprotein, a small--presumably
non-virion--protein, and a polymerase protein. The presence of a non-virion protein,
which is the hallmark feature of all Novirhabdoviruses, supported SHRV's identity as a
member of the Novirhabdovirus genus, despite the fact that the non-virion protein
showed no homology with any known protein.
A system was developed to express a full-length, error-free positive-strand copy
of SHRV's RNA genome along with all of the SHRV proteins required for viral
replication within the cytoplasm of a virus-susceptible host cell. These factors
collectively allowed the recovery of live virus entirely from cloned cDNAs. A unique
restriction site was engineered into SHRV's cDNA genome, and the presence of this
restriction site was verified following virus recovery, proving the recovered virus was
indeed a live recombinant virus. To our knowledge this achievement marks the first time
in which reverse genetics has been performed on a nonmammalian negative-stranded
RNA virus. / Graduation date: 1999
| Identifer | oai:union.ndltd.org:ORGSU/oai:ir.library.oregonstate.edu:1957/33875 |
| Date | 25 February 1999 |
| Creators | Johnson, Marc C. |
| Contributors | Leong, Jo-Ann C. |
| Source Sets | Oregon State University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
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