History of mycoplasma strains and plasmid
screening. (XLS 32 KB) 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). ARN-509 order (XLSX 22 KB) 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 this website 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 Blasticidin S molecular weight initiation
site. The box CAT indicates the initiation codon of the rep gene that is encoded on the complementary DNA strand. (DOCX 37 KB) Additional file 5: Figure S2. Detection of pMyBK1 ssDNA intermediates before 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. (PPTX 122 KB) 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. (PPTX 85 KB) References 1. Smets BF, Barkay T: Horizontal gene transfer: perspectives at a crossroads of scientific disciplines. Nature Reviews 2005,3(9):675–678.PubMedCrossRef 2. Frost LS, Leplae R, Summers AO, Toussaint A: Mobile genetic elements: the agents of open source evolution. Nature reviews 2005,3(9):722–732.PubMedCrossRef 3. Razin S, Yogev D, Naot Y: Molecular biology and pathogenicity of mycoplasmas. Microbiol Mol Biol Rev 1998,62(4):1094–1156.PubMed 4.