Rubella virus is a human pathogen which can cause birth defects in pregnant women and long term virus persistence in some individuals. The implications of rubella persistence are incompletely understood. However, rubella persistence has been linked to vaccine failure, some forms of chronic arthritis and a rare, fatal panencephalitis years after primary infection. The three major goals of the proposed research are. first, to finish the nucleotide sequencing of the rubella genomic RNA. At the time of writing, we have determined the sequence of the 3' terminal 2,400 nucleotides of the rubella genome. The rubella genome is a single stranded, positive-polarity RNA of roughly 10,000 nucleotides. The 3500 3' nucleotides contain the coding regions for the virus structural proteins while the 5' 6500 nucleotides contain the coding region for several nonstructural proteins which have not been identified. To identify these nonstructural proteins, the deduced nonstructural protein amino acid sequence will be used as a guide for the synthesis of peptides against which antibodies will be raised and used to immunoprecipitate the nonstructural proteins from extracts of infected cells. The second major goal is to introduce the coding regions for the virus structural proteins into Vaccinia virus. Vaccinia virus recombinants containing the rubella structural proteins are potential vaccines and will be tested for ability to induce production of neutralizing antibodies in mice. Manipulation of the rubella structural proteins in a Vaccinia vector will also lead to development of reagents for detection and study of the humoral and cell mediated immune response against rubella. Thirdly, the extent of the rubella genome which has been introduced into a plasmid containing the promoters for SP6 and T7 RNA polymerases will be increased substantially (currently, the 3' 600 nucleotides of the genome have been cloned into such a vector). By so doing, the sensitivity of RNA hybridization probes synthesized from these plasmids will be increased significantly. The current plasmid has been distributed to four investigators who are attempting to detect the presence of rubella RNA by in situ hybridization in arthritis biopsies, peripheral blood cells from congenital rubella patients, and brain autopsies from progressive rubella panencephalitis patients.. The expanded plasmid will likewise be made available to investigators doing this type of research.
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