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Journal of Bacteriology, April 2001, p. 2157-2164, Vol. 183, No. 7
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.7.2157-2164.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Genomic Sequence and Transcriptional Analysis of a 23-Kilobase Mycobacterial Linear Plasmid: Evidence for Horizontal Transfer and Identification of Plasmid Maintenance Systems

Corinne Le Dantec,1 Nathalie Winter,2 Brigitte Gicquel,3 Véronique Vincent,1 and Mathieu Picardeau4,*

Laboratoire de Référence des Mycobactéries,1 Laboratoire du B.C.G.,2 Unité de Génétique Mycobactérienne,3 and Unité de Bactériologie Moléculaire et Médicale,4 Institut Pasteur, 75724 Paris Cedex 15, France

Received 16 October 2000/Accepted 8 January 2001

Linear plasmids were unknown in mycobacteria until recently. Here, we report the complete nucleotide sequence of 23-kb linear plasmid pCLP from Mycobacterium celatum, an opportunistic pathogen. The sequence of pCLP revealed at least 19 putative open reading frames (ORFs). Expression of pCLP genes in exponential-phase cultures was determined by reverse transcriptase PCR (RT-PCR). Twelve ORFs were expressed, whereas no transcription of the 7 other ORFs of pCLP was detected. Five of the 12 transcribed ORFs detected by RT-PCR are of unknown function. Sequence analysis revealed similar loci in both M. celatum pCLP and the Mycobacterium tuberculosis chromosome, including transposase-related sequences. This result suggests horizontal transfer between these two organisms. pCLP also contains ORFs that are similar to genes of bacterial circular plasmids involved in partition (par operon) and postsegregational (pem operon) mechanisms. Functional analysis of these ORFs suggests that they probably carry out similar maintenance roles in pCLP.

* Corresponding author. Mailing address: Unité de Bactériologie Moléculaire et Médicale, 28 rue du Dr Roux, Institut Pasteur, 75724 Paris Cedex 15, France. Phone: (33) 1 45 68 80 00, ext. 7233. Fax: (33) 1 40 61 30 01. E-mail: mpicard{at}pasteur.fr.

Journal of Bacteriology, April 2001, p. 2157-2164, Vol. 183, No. 7
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.7.2157-2164.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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