Abstract
Mycobacteria, like other bacteria, respond to environmental signals with changes in gene expression regulated at the level of transcription. The significance of gene-expression patterns in pathogens, such asMycobacterium tuberculosis, is that specific signals encountered durmg the process of mfection regulate, often via two component regulatory systems, the expression of vn-ulence genes necessary for that stage of host-pathogen mteraction. Signals known to Induce bacterial gene expression include temperature, osmolarity, oxidative stress, pH, low oxygen tension, nutrient deprivation, and Fe2+ limitation different conditions inducing different, but overlapping subsets of genes (1), sometimes referred to as stimulons, regulons, or virulons. Little is known about gene-expression patterns of mycobacteria and the definition of differentially -expressed genes is central to understanding pathogenesis and virulence at a molecular level.
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References
Mekalanos, J J. (1992) Environmental signals controllmg expression of vn-ulence determinants in bacterta. J Bacteriol 174, 1–7
Wren, B. W., Colby, S. M, Cubberley, R. R., and Pallen, M J (1992) Degenerate primers for the amplificatton of fragments from genes encoding response regulators from a range of pathogenic bacteria FEMS Mlcroblol Lett 99, 287–292
Sherman, D R, Sabo, P J., Hockey, M J, Aram, T. M., Mahanas, G. G, Yuan, Y, Barr, III, C E, and Stover, C K (1995) Disparate responses to oxidative stress in saprophytrc and pathogemc mycobactena. Proc Natl Acad Sci USA 92, 6625–6629
Patel, B. K. R., BanerJee, D. K., and Butcher, P. D. (1991) Charactertsation of the heat shock response in M bows BCG. J Bacterlol 173, 7982–7987
Lathtgra, R. B., Butcher, P. D., Garbe, T. R., and Young, D. B. (1991) Heat shock proteins as virulence factors of pathogens Curr Top. Mlcroblol Immunol. 167, 125–143
Garbe, T. R., Hibler, N S, and Deretlc, V. (1996) Response of Mycobacterzum tuberculosis to reactive oxygen intermediates. Mol Med 2, 134–142
Plum, G. and Clark-Curtis, J E. (1993) Induction of Mycobacterium avzum gene expresslon following phagocytosls by human macrophages Infect Immun 62, 476–483.
Alavl, M. R and Affronti L F (1994) Induction of mycobacterial proteins durmg phagocytosls and heat shock. a hme interval analysis J. Leukoc BloI 55, 633–641
Lee, B.-Y. and Horwitz, M A (1995) Identtficatlon of macrophage and stress induced proteins of Mycobacterium tuberculosis. J. Clin Invest. 96, 245–249.
Dellagostin, 0 A., Esposito, G., Eales, L.-J., Dale, J W, and McFadden, J J (1995) Activity of mycobacterlal promotors during intracellular and extracellular growth. Microbiology 141, 1785–1792.
Llsltsyn, N., Llsltsyn, N., and Wigler, M. (1993) Cloning the difference between two complex genomes. Science 259, 946–951
Hubank, M. and Schatz, D. G. (1994) Identifying differences in mRNA expression by representational difference analysis of cDNA Nucleic Acids Res 22, 5640–5648
Kmger, A. K. and Tyagi, J. S (1993) Identification and clomng of genes dlfferentlally expressed in the virulent strian of Mycobacterium tuberculosis Gene 131, 113–117.
Liang, P. and Pardee, A B. (1992) Differential display of eukaryotic mRNA by means of the polymerase chain reaction Science 257, 967–971
Welsh, J, Chada, S., Dalal, S, Cheng, R, Ralph, D., and McClelland, M (1992) Arbitrarily primed PCR fingerprinting of RNA Nucleic Acrds Res. 20, 4965–4970.
Manley, J. L. (1995) Messenger RNA polyadenylatlon. a universal modrficatlon Proc Natl Acad Sci USA 92, 1800–1801
Wong, K. K and McClelland, M. (1994) Stress-inducible gene of SaEmoneiZa typhtmurlum identified by arbitrarily primed PCR of RNA Proc Natl Acad Sci USA 91, 639–643.
Patel, B. K. R., Baneqee, D. K., and Butcher, P. D. (1991) Extraction and characterisation of mRNA from mycobactena: lmplicatlon for virulence gene ldentlfication J, Mlcrobiol Methods 13, 99–111.
Froussard, P. (1992) A random-PCR method to construct whole cDNA hbrary from low amounts of RNA Nuclezc Acids Res 20, 290O.
Ralph, D., McClelland, M., and Welsh, J. (1993) RNA fingerprinting using arbltrarlly primed PCR identifies differentially regulated RNAs in mink lung (My 1 Lu) cells growth arrested by transforming growth factor beta 1. Proc Natl Acad Scz. USA 90, 10,710–10,714
Lohmann, J., Schickle, H., and Bosch, T. C. G. (1995) REN display, a rapid and efficient method for nonradioactive differential display and mRNA lsolatlon BzoTechnzques l8, 200–202.
Bassam, B. J, Caetano-Annolles, G., and Gresshoff, P. M. (1991) Fast and sensetive stlver staining of DNA in polyacrylamlde gels. Anal Bzochem 196, 80–83
Bauer, D, Muller, H, Reich, J, Riedel, H., Ahrenkle, V., Warthoe, P., and Strauss, M.(1993) Identification of differentially expressed mRNA species by an improved display technrque (DDRT-PCR) Nuclezc Ads Res 21, 4272–4280.
Kim, A, Roffler-Tarlov, S, and Lin, C S (1995) New technique for precise ahgnment of an RNA differential display gel with its film image BzoTechnzques 19, 346
Dlachenko, L. B, Ledesma, J, Chenchik, A A., and Siebert, P D (1996) Combining the techmque of RNA fingerprmtmg and differential display to obtain dlfferentially expressed mRNA. Bzochem. Bzophys. Res Comm. 219, 824–828.
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© 1998 Humana Press Inc., Totowa, NJ
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Mangan, J.A., Butcher, P.D. (1998). 24 Analysis of Mycobacterial Differential Gene Expression by RAP-PCR. In: Parish, T., Stoker, N.G. (eds) Mycobacteria Protocols. Methods in Molecular Biology™, vol 101. Humana Press. https://doi.org/10.1385/0-89603-471-2:307
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DOI: https://doi.org/10.1385/0-89603-471-2:307
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