Our long-term goal is to understand basic processes involved in animal virus gene expression, including transcription, translation, and assembly of viral membranes. In addition, we are interested in basic questions about the potential usefulness of recombinant DNA technology in production of anti-viral vaccines. Four major areas of research are contemlated: We plan additional studies on transcription of the vesicular stomatitis virus (VSV) genome with emphasis on understanding the mechanisms which generate sequential mRNA transcription and localized transcription attenuation. We will continue our analysis of ribosome recognition of viral mRNA by determining if two """"""""extra"""""""" ribosome binding sites in a single viral mRNA species are actually used in vivo. We plan to use the VSV glycoprotein as a model system for understanding the assembly of viral and cellular membranes. These studies will focus on determination of protein sequence changes in mutant G proteins which are blocked at different stages of intracellular transport, and on the sequences of glycoproteins from different serotypes. Sequences will be determined from primers extended on the genomic RNA derived from G mutants. We will continue our studies on expression of the VSV glycoprotein in E. coli to determine if high level expression of the protein is possible, and if the protein produced is capable of inducing neutralizing antibody and immunity to viral infection in animals.
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