Open Access Research article

Distribution and diversity of mycoplasma plasmids: lessons from cryptic genetic elements

Marc Breton12, Florence Tardy3, Emilie Dordet-Frisoni45, Eveline Sagne45, Virginie Mick3, Joël Renaudin12, Pascal Sirand-Pugnet12, Christine Citti45 and Alain Blanchard126*

Author Affiliations

1 University Bordeaux, UMR 1332 Biologie du Fruit et Pathologie, 71 avenue Edouard Bourlaux, F-33140, Villenave d'Ornon, France

2 INRA, UMR 1332 Biologie du Fruit et Pathologie, 71, avenue Edouard Bourlaux, F-33140, Villenave d'Ornon, France

3 Anses, Laboratoire de Lyon, UMR Mycoplasmoses des Ruminants, 31 Avenue Tony Garnier, F-69364, Lyon cedex 07, France

4 INRA, UMR1225, Ecole Nationale Vétérinaire de Toulouse, 23 Chemin des Capelles, F-31076, Toulouse Cedex 3, France

5 Université de Toulouse, INP-ENVT, UMR1225, Ecole Nationale Vétérinaire de Toulouse, 23 Chemin des Capelles, F-31076, Toulouse Cedex 3, France

6 Centre INRA de Bordeaux Aquitaine, UMR 1332 Biologie du Fruit et Pathologie, 71, avenue Edouard Bourlaux, BP81, F-33140, Villenave d'Ornon, France

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BMC Microbiology 2012, 12:257  doi:10.1186/1471-2180-12-257

Published: 12 November 2012

Additional files

Additional file 1:

Table S1. Additional file 5.

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Additional file 2:

Table S2. History of mycoplasma strains and plasmid screening.

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Additional file 3:

Table S3. Pairwise nucleic sequence identities between mycoplasma plasmids. Global alignments of the full-length nucleic sequence of mycoplasma plasmids were accomplished using a Needleman–Wunsch algorithm implemented in the Needleall program (Needleman & Wunsch, 1970). Identity percents are indicated. Rep group refers to Rep phylogeny (see Figure 6). Table S3. Pairwise nucleic sequence identities between mycoplasma plasmids. Global alignments of the full-length nucleic sequence of mycoplasma plasmids were accomplished using a Needleman–Wunsch algorithm implemented in the Needleall program (Needleman & Wunsch, J Mol Biol 1970;48:443-53). Identity percents are indicated. Rep group refers to Rep phylogeny (see Figure 6).

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Additional file 4:

Figure S1. Nucleotide sequences of the predicted ctRNA coding strands. The counter-transcripts were first identified by analogy with those of pMV158 or its derivative pLS1. These ctRNA overlap the rep gene start and have a length of only a few tens of nucleotides. Using the consensus sequence TTGACA – (N17) –TG-N-TATAAT for the promoter, putative promoters were identified in the aligned sequences. Putative Pct promoters are indicated with the -35 and -10 regions in bold and underlined letters. Arrows indicate inverted repeats of the putative rho independent terminators. The ctRNA of pLS1 (rnaII) is shown as proposed by del Solar et al. [46] with an arrowhead indicating the possible transcriptional initiation site. The box CAT indicates the initiation codon of the rep gene that is encoded on the complementary DNA strand.

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Additional file 5:

Figure S2. Detection of pMyBK1 ssDNA intermediates by Southern blot hybridization. Total DNA from Mycoplasma yeatsii type strain GIH TS (lane 1-2) was analyzed on a 0.8% agarose gel (A) with (+) or without (-) prior S1 nuclease treatment. Southern blot (B) was performed with digoxigenin-labeled pMyBK1 probe under non-denaturing conditions. M, DNA ladder.

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Additional file 6:

Figure S3. Expression of spiralin in Mcc using pMyBK1 derivatives. Whole cell dot immunoblot of 12 Mcc transformants harboring the spiralin expression vector pCM-K3-spi (a) or the empty vector pCM-K3 (b). Mycoplasma cells were applied to a nitrocellulose membrane and probed with rabbit anti-spiralin antibodies and anti-rabbit IgG peroxidase conjugate.

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