The goal is to study regulation of ribosome synthesis in cultured mosquito cells and in mosquito fat body during a vitellogenic cycle. Specifically, we are interested in using gene probes derived from work with cultured cells, together with DNA-mediated gene transfer technology, to understand in greater detail regulatory mechanisms that affect the protein synthetic capability of insect cells. We plan to substantiate, by protein chemistry, the identity of cDNA clones corresponding to ribosomal proteins, L8, L14, and L31, and use these cDNA clones (corresponding to ribosomal protein mRNAs), to isolate from a genomic library the corresponding genes. These genes will be characterized to identify exon/intron organization, copy number, and regulatory sequences. Ribosome synthesis and stability will be measured in mosquito fat body. Gene probes will be used to assay ribosomal protein mRNA synthesis in fat body during the vitellogenic cycle. Transcriptional and/or translational regulatory processes will be identified. Gene transfer technology will be studies in further detail. Methods that have been worked out using transient expression assays will be adapted to achieve stable gene transfer and to accomplish plasmid rescue of a selectable marker. Attempts will be made to extend DNA-mediated gene transfer methodology to the intact mosquito. This work will provide basic information on the biochemistry and physiology of the mosquito, a disease vector of considerable economic importance.
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