Genetically Engineered Bacteria to Identify and Produce Anti-Viral Agents
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We have prepared a strain of Escherichia coli that expresses both the HIV protease and a Tet protein which has been modified to contain the HIV protease recognition sequence. When the protease is expressed, the bacteria will not grow in the presence of tetracycline. However, when the protease is inhibited the bacteria can grow in tetracycline containing media (Block and Grafstrom 1990). We have selected spontaneously arising Tet resistant mutants and have screened them for those that could be producing an inhibitor of HIV protease. The problems in the construction of this strain and the characterization of the various Teti mutants are discussed.
KeywordsHuman Immunodeficiency Virus Antiviral Agent Tetracycline Resistance Tetracycline Resistance Gene Mutant Bacterium
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- Borel, J.F., Feurer, C., Magnee, C., and Stahelin, H., 1977, Effects of the new anti-lymphocytic peptide cyclosporin A in animals, Immunol. 32:1017–1025.Google Scholar
- Debouck, C., Gorniak, J.G., Strickler, J.E., Meek, T.D., Metcalf, B.W., and Rosenberg, M., 1987, Human immunodeficiency virus protease expressed in Escherichia coli exhibits autoprocessing and specific maturation of the gag precursor, Proc. Natl. Acad.Sci. USA 84:8903–8906.PubMedCrossRefGoogle Scholar
- DeClercq, KE., 1990, Targets and strategies for the antiviral chemotherapy of AIDS, TIPS 11:198–205.Google Scholar
- Dreyer G.B., Metcalf, B.W., Tomaszek, T.A., Jr., Carr, T.J., Chandler, A.C., III, Hyland, L., Fakhouryk, S.A., Magnard, V.W., Moore, M.L., Strickler, J.F., Debouck, C., and Meek, T.D.1989, Inhibition of HIV-1 protease in vitro: rational design of substrate analogue inhibitors, Proc. Natl.Acad. Sci. USA 86:9752–9756.PubMedCrossRefGoogle Scholar
- Kholberg, R., 1991, Critics call for a smarter way to screen for drugs, J. NIH Res. 3:25–26.Google Scholar
- Korant, B., 1990, AIDS Research and Reference Reagent Program Catalog, U.S. Department of Health and Human Services, p.51 (January).Google Scholar
- Legrice, S.F.J., Mills, J., and Mous, J., 1988, Active site mutagenesis of the AIDS virus protease and its alleviation by trans complementation, EMBO J. 7:2547–2553.Google Scholar
- Moore, M.L., Bryan, W.M., Fakhoury, S.A., and Maagard, V.M., Huffman, W.F., Dayton, B.D., Meek, T.D., Hyland, L., Dreyer, G.B., Metcalf, B.W., Strickler, J.E., Gorniak, J.G., and Debouck, C., 1989, Peptide substrates and inhibitors of the HIV-1 protease, Biochem. Biophys. Res. Comm. 159:420–425.PubMedCrossRefGoogle Scholar
- Pauwels, R., Andries, K., Desmyter, J., Schols, D., Kukla, M.J., Breslin, H.J., Raeymaeckers, A., Van Gelder, J., Woestenborghs, R., Heykants, J., Schellekens, K., Janssen, M.A.C., DeClerq, E., and Janssen, P.A.J., 1990, Potent and selective inhibition of HIV-1 replication in vitro by a novel series of TIBO derivatives, Nature 343:470–474.PubMedCrossRefGoogle Scholar
- Roberts, N.A., Martin, J.A., Kinchington, D., Broadhurst, A.V., Craig, J.C., Duncan, I.B., Galpin, S.A., Handa, B.K., Krohn, J.K.A., Lambert, R.W., Merrett, J.H., Mills, J.S., Parkes, K.B.B., Redshaw, S., Ritchie, A.J., Taylor, D.L., Thomas, G.J., Machin, P.J., 1990, Rational design of peptide-based HIV proteinase inhibitors, Science 248:358–361.PubMedCrossRefGoogle Scholar
- Sidwell, R.W., Dixon, G.J., Schabel, F.M., 1968, Antiviral activity of 9-beta-Darabinofuranosyl adenine, Antimicrob. Agent. Chemo. 8:148–154.Google Scholar