Abstract
Prokaryotic insertion sequences (IS) are transposable genetic elements with a length ranging from 800–2500 bp. They are found at a remarkable variety in the genomes of many different bacteria and mycoplasmas at a multiplicity of between a few and several hundred per genome. In bacteria, they are very frequently found as part of natural plasmids. Genetic phenomena associated with transposition of IS elements are: spontaneous, often highly polar mutations, sometimes associated with deletion or inversion of adjacent DNA segments, as well as activation of the transcription of flanking genes (1). Insertion sequences therefore contribute basically to the variability of the prokaryotic genome and phenotype, and are thought to play an important role in the adaptability of prokaryotes to the environment. Insertion sequences provide a valuable source of experimental material for the study of gene expression, recombination and repair events, population dynamics, and horizontal transmission of genes. They are also frequently used in molecular genetics as markers to facilitate gene mapping, as mutagens, and for gene cloning. Several insertion sequences have been used as markers for molecular DNA fingerprints for identification and subtyping of bacterial and mycoplasmal strains, such as Mycobacterium tuberculosis (2,3), Salmonella typhimurium (4), Vibrio cholerae (5), and Mycoplasma mycoides subsp. mycoides small-colony type (SC) (6).
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References
Galas, D. J. and Chandler, M. (1989) Bacterial insertion sequences. Mobile DNA (Berg, D. E. and Howe, M. M., eds.), American Society for Microbiology, Washington, DC, pp. 109–162.
van Embden, J. D., Cave, M. D., Crawford, J. T., Dale, J. W., Eisenach, K. D., Gicquel, et al. (1993) Strain identification of Mycobacterium tuberculosis by DNA fingerprinting: recommendations for a standardized methodology. J. Clin. Microbiol. 31, 406–409.
Small, P. M., Hopewell, P. C., Singh, S. P.; Paz, A., Parsonnet, J.; Ruston, D. C., et al. (1994) The epidemiology of tuberculosis in San Francisco. A population-based study using conventional and molecular methods. N. Engl. J. Med. 330, 1703–1709.
Stanley, J., Baquar, N., and Threlfall, E. J. (1993) Genotypes and phylogenetic relationships of Salmonella typhimurium are defined by molecular fingerprinting of IS200 and 16S rrn loci. J. Gen. Microbiol. 139, 1133–1140.
Bik, E. M., Gouw, R. D., and Mooi, F. R. (1996) DNA fingerprinting of Vibrio cholerae strains with a novel insertion sequence element-a tool to identify epidemic strains. J. Clin. Microbiol. 34, 1453–1461.
Cheng, X. X., Nicolet, J., Poumarat, F.; Regalla, J., Thiaucourt, F., and Frey, J. (1995) Insertion element IS1296 in Mycoplasma mycoides subsp. mycoides small colony identifies a European clonal line distinct from African and Australian strains. Microbiology 141, 3221–3228.
Frey, J., Cheng, X. X., Kuhnert, P., and Nicolet, J. (1995) Identification and characterization of IS1296 in Mycoplasma mycoides subsp. mycoides SC and presence in related mycoplasmas. Gene 160, 95–100.
Bhugra, B. and Dybvig, K. (1993) Identification and characterization of IS1138, a transposable element from Mycoplasma pulmonis that belongs to the IS3 family. Mol. Microbiol. 7, 577–584.
Ferrell, R. V., Heidari, M. B., Wise, K. S., and McIntosh, M. A. (1989) A Mycoplasma genetic element resembling prokaryotic insertion sequences. Mol. Microbiol. 3, 957–967.
Hu, W. S., Wang, R. Y., Liou, R. S., Shih, J. W., and Lo, S. C. (1990) Identification of an insertion-sequence-like genetic element in the newly recognized human pathogen Mycoplasma incognitus. Gene 93, 67–72.
Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A., et al. (1990) Current Protocols in Molecular Biology. Wiley Interscience, New York.
Pitcher, D. G., Saunders, N. A., and Owen, R. J. (1989) Rapid extraction of bacterial genomic DNA with guanidium thiocyanate. Lett. Appl. Microbiol. 8, 151–156.
Sneath, P. H. A. and Sokal, R. R. (1973) Numerical Taxonomy. The Principles and Practice of Numerical Classification. W. H. Freeman, San Francisco.
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© 1998 Humana Press Inc., Totowa, NJ
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Frey, J. (1998). Insertion Sequence Analysis. In: Miles, R., Nicholas, R. (eds) Mycoplasma Protocols. Methods in Molecular Biology™, vol 104. Humana Press. https://doi.org/10.1385/0-89603-525-5:197
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DOI: https://doi.org/10.1385/0-89603-525-5:197
Publisher Name: Humana Press
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