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Journal of Bacteriology, September 1999, p. 5711-5717, Vol. 181, No. 18
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Analysis of Fusion Junctions of Circularized Chromosomes in Streptomyces griseus

Daisuke Kameoka, Alexander Lezhava, Hiroyuki Zenitani, Keiichiro Hiratsu, Makoto Kawamoto, Kohei Goshi, Kuninobu Inada, Hidenori Shinkawa, and Haruyasu Kinashi*

Department of Molecular Biotechnology, Hiroshima University, 1-4-1 Kagamiyama, Higashi-Hiroshima 739-8527, Japan

Received 5 February 1999/Accepted 30 May 1999

A filamentous soil bacterium, Streptomyces griseus 2247, carries a 7.8-Mb linear chromosome. We previously showed by macrorestriction analysis that mutagenic treatments easily caused deletions at both ends of its linear chromosome and changed the chromosome to a circular form. In this study, we confirmed chromosomal circularization by cloning and sequencing the junction fragments from two deletion mutants, 404-23 and N2. The junction sequences were compared with the corresponding right and left deletion end sequences in the parent strain, 2247. No homology and a 6-bp microhomology were found between the two deletion ends of the 404-23 and N2 mutants, respectively, which indicate that the chromosomal circularization was caused by illegitimate recombination without concomitant amplification. The circularized chromosomes were stably maintained in both mutants. Therefore, the chromosomal circularization might have occurred to prevent lethal deletions, which otherwise would progress into the indispensable central regions of the chromosome.

* Corresponding author. Mailing address: Department of Molecular Biotechnology, Hiroshima University, 1-4-1 Kagamiyama, Higashi-Hiroshima 739-8527, Japan. Phone and fax: 81 (824) 24 7869. E-mail: kinashi{at}ipc.hiroshima-u.ac.jp.

Journal of Bacteriology, September 1999, p. 5711-5717, Vol. 181, No. 18
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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