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J Bacteriol, April 1998, p. 2102-2109, Vol. 180, No. 8
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

IS6110 Transposition and Evolutionary Scenario of the Direct Repeat Locus in a Group of Closely Related Mycobacterium tuberculosis Strains

Z. Fang,1 N. Morrison,1 B. Watt,2 C. Doig,2 and K. J. Forbes1,*

Medical Microbiology, Aberdeen University, Foresterhill, Aberdeen, AB25 2ZD,1 and Scottish Mycobacteria Reference Laboratory, City Hospital, Edinburgh, EH10 5SB,2 United Kingdom

Received 11 August 1997/Accepted 6 February 1998

In recent years, various polymorphic loci and multicopy insertion elements have been discovered in the Mycobacterium tuberculosis genome, such as the direct repeat (DR) locus, the major polymorphic tandem repeats, the polymorphic GC-rich repetitive sequence, IS6110, and IS1081. These, especially IS6110 and the DR locus, have been widely used as genetic markers to differentiate M. tuberculosis isolates and will continue to be so used, due to the conserved nature of the genome of M. tuberculosis. However, little is known about the processes involved in generating these or of their relative rates of change. Without an understanding of the biological characteristics of these genetic markers, it is difficult to use them to their full extent for understanding the population genetics and epidemiology of M. tuberculosis. To address these points, we identified a cluster of 7 isolates in a collection of 101 clinical isolates and investigated them with various polymorphic genetic markers, which indicated that they were highly related to each other. This cluster provided a model system for the study of IS6110 transposition, evolution at the DR locus, and the effects of these on the determination of evolutionary relationships among M. tuberculosis strains. Our results suggest that IS6110 restriction fragment length polymorphism patterns are useful in grouping closely related isolates together; however, they can be misleading if used for making inferences about the evolutionary relationships between closely related isolates. DNA sequence analysis of the DR loci of these isolates revealed an evolutionary scenario, which, complemented with the information from IS6110, allowed a reconstruction of the evolutionary steps and relationships among these closely related isolates. Loss of the IS6110 copy in the DR locus was noted, and the mechanisms of this loss are discussed.

* Corresponding author. Mailing address: Medical Microbiology, Aberdeen University, Foresterhill, Aberdeen, AB25 2ZD, United Kingdom. Phone: 44 1224 663123, ext. 54953. Fax: 44 1224 685604. E-mail: k.forbes{at}abdn.ac.uk.




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