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Applied and Environmental Microbiology, May 2006, p. 3206-3216, Vol. 72, No. 5
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.5.3206-3216.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Characterization of the Replication, Maintenance, and Transfer Features of the IncP-7 Plasmid pCAR1, Which Carries Genes Involved in Carbazole and Dioxin Degradation{dagger}

Masaki Shintani,1 Hirokazu Yano,2 Hiroshi Habe,1 Toshio Omori,3 Hisakazu Yamane,1 Masataka Tsuda,2 and Hideaki Nojiri1*

Biotechnology Research Center, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657,1 Department of Environmental Life Sciences, Graduate School of Life Sciences, Tohoku University, 2-1-1 Katahira, Sendai 980-8577,2 Department of Industrial Chemistry, Shibaura Institute of Technology, 3-9-14 Shibaura, Minato-ku, Tokyo 108-8548 Japan3

Received 25 December 2005/ Accepted 8 February 2006

Isolated from Pseudomonas resinovorans CA10, pCAR1 is a 199-kb plasmid that carries genes involved in the degradation of carbazole and dioxin. The nucleotide sequence of pCAR1 has been determined previously. In this study, we characterized pCAR1 in terms of its replication, maintenance, and conjugation. By constructing miniplasmids of pCAR1 and testing their establishment in Pseudomonas putida DS1, we show that pCAR1 replication is due to the repA gene and its upstream DNA region. The repA gene and putative oriV region could be separated in P. putida DS1, and the oriV region was determined to be located within the 345-bp region between the repA and parW genes. Incompatibility testing using the minireplicon of pCAR1 and IncP plasmids indicated that pCAR1 belongs to the IncP-7 group. Monitoring of the maintenance properties of serial miniplasmids in nonselective medium, and mutation and complementation analyses of the parWABC genes, showed that the stability of pCAR1 is attributable to the products of the parWAB genes. In mating assays, the transfer of pCAR1 from CA10 was detected in a CA10 derivative that was cured of pCAR1 (CA10dm4) and in P. putida KT2440 at frequencies of 3 x 10–1 and 3 x 10–3 per donor strain, respectively. This is the first report of the characterization of this completely sequenced IncP-7 plasmid.

* Corresponding author. Mailing address: Biotechnology Research Center, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. Phone: 81-3-5841-3064. Fax: 81-3-5841-8030. E-mail: anojiri{at}mail.ecc.u-tokyo.ac.jp.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, May 2006, p. 3206-3216, Vol. 72, No. 5
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.5.3206-3216.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Takahashi, Y., Shintani, M., Li, L., Yamane, H., Nojiri, H. (2009). Carbazole-Degradative IncP-7 Plasmid pCAR1.2 Is Structurally Unstable in Pseudomonas fluorescens Pf0-1, Which Accumulates Catechol, the Intermediate of the Carbazole Degradation Pathway. Appl. Environ. Microbiol. 75: 3920-3929 [Abstract] [Full Text]  
  • Miyakoshi, M., Shintani, M., Terabayashi, T., Kai, S., Yamane, H., Nojiri, H. (2007). Transcriptome Analysis of Pseudomonas putida KT2440 Harboring the Completely Sequenced IncP-7 Plasmid pCAR1. J. Bacteriol. 189: 6849-6860 [Abstract] [Full Text]  
  • Sota, M., Tsuda, M., Yano, H., Suzuki, H., Forney, L. J., Top, E. M. (2007). Region-Specific Insertion of Transposons in Combination with Selection for High Plasmid Transferability and Stability Accounts for the Structural Similarity of IncP-1 Plasmids. J. Bacteriol. 189: 3091-3098 [Abstract] [Full Text]  
  • Shintani, M., Urata, M., Inoue, K., Eto, K., Habe, H., Omori, T., Yamane, H., Nojiri, H. (2007). The Sphingomonas Plasmid pCAR3 Is Involved in Complete Mineralization of Carbazole. J. Bacteriol. 189: 2007-2020 [Abstract] [Full Text]  


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