The goal of the DNA SEQUENCING CORE is to: (1) provide a core laboratory for high through put DNA sequencing of genes, compilation of data and analysis of the edited DNA sequences contributed by the center's investigators; (2) provide for the construction of expression plasmids and recombinant replication competent pro-viruses and vaccinia virus vectors; and (3) provide computer-based analysis of viral RNA secondary structure using the combined expertise available at the University of Wisconsin through the Genetics Computer Group and the super-computer facility at the University of Minnesota to analyze the secondary structures of HIV-1 mutants and vectors. The DNA sequencing core will facilitate interactions among researchers, allow researchers to readily access DNA sequencing and analysis and molecular reconstructions, lower the cost of this technology to individual researchers, expedite RNA structural analysis and facilitate the undertaking of new projects by aiding in vector design and DNA sequencing analysis. The DNA sequencing core will perform high through put DNA sequencing of genes or portions of genes, compile data and analyze the edited DNA sequences contributed by the investigators of the center. Additionally, this core will construct expression plasmids, recombinant pro-viruses and recombinant vaccinia virus vectors. Dr. Wolinsky, the core director, and Dr. Clark Brown and Ms. Zareefa Burki, his staff at Northwestern University will be responsible for bi-directional DNA sequencing using a dedicated ABI 377 automated sequencer, sequence analysis using UNIX-based sequence analysis algorithms, and vector construction. RNA structure analysis using state-of-the-art computational techniques to generate and evaluate candidate RNA structures composed of local nucleotides as well as long range interactions that impact on the higher order organization of the viral RNA will complement nucleic acid sequence analysis. Dr. Antonito Panganiban of the University of Wisconsin will direct this component of the core.
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