Molecular characterization and genetic recombination of snakehead rhabdovirus Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/mg74qp645

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  • 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.
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  • description.provenance : Approved for entry into archive by Patricia Black(patricia.black@oregonstate.edu) on 2012-09-25T21:09:49Z (GMT) No. of bitstreams: 1 JohnsonMarcC1999.pdf: 7938273 bytes, checksum: bec805c69894699e00750f94f22c2049 (MD5)
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