Analysis of transconjugants isolated in mating experiments using <i>M. avium</i> 88.3 as donor strain.

<p>(A) PFGE with undigested DNA; (B) Southern blot of PFGE gels with undigested DNA and hybridization with IS<i>1245</i>-derived probe; (C) PFGE-DraI; (D) Southern blot of PFGE-DraI gels and hybridization with pMA100-derived probe. Open arrows indicate the linear plasmid pMA100. 1: <i>M. avium</i> 88.3; 2: <i>M. kansasii</i> 88.6; 3: <i>M. kansasii</i> IAL 413; 4: <i>M. kansasii</i> IEC 6805; 5: <i>M. bovis</i> BCG Moreau; W: wild-type colony; T: transconjugant colony.</p

PFGE and Southern blot hybridization with IS<i>1245</i>-derived probe of

<p><b><i>M. avium</i></b><b> and </b><b><i>M. kansasii</i></b><b> colonies.</b> (A) PFGE with undigested DNA; (B) Southern blot hybridization with IS<i>1245</i>-derived probe. Open arrow indicates pMA100; closed arrow indicates the uncharacterized smaller hybridization band. 88.1 to 88.4 = <i>M</i>. <i>avium</i>; 88.5 to 88.75 = PCR−IS<i>1245-</i>negative <i>M</i>. <i>kansasii</i>; 88.8 to 88.15 = PCR−IS<i>1245-</i>positive <i>M. kansasii.</i> On the left, Lambda Ladder PFG Marker (NewEngland BioLabs) molecular size markers.</p

Conjugation experiments using <i>M. avium</i> 88.3 as the donor strain.

<p>*three experiments with 15, 20 and 25 days incubations, respectively.</p><p>**three experiments with 2, 4 and 10 days incubations, respectively.</p><p>§pigmented colonies after exposure to light.</p><p>£PCR-IS<i>6110</i> positive.</p><p>¶grown on LB agar.</p

PFGE of DNA genomic preparations.

<p>(A) PFGE with undigested DNAs from <i>M. avium</i> 88.3 (1) and <i>M. kansasii</i> 88.8 (2) under different switch times, indicated below each figure; (B) pMA100 extracted from PFGE gels and treated with exonuclease III (3) or exonuclease lambda (4); (C) pMA100 extracted from PFGE gels and treated (+) or not (-) with topoisomerase I; (D) DNA prepared with (+) or without (-) adding proteinase K to the lysis buffer; (E) same as in (D) in PFGE gels and running buffer prepared with 0.2% SDS. λ: DNA concatemers of the bacteriophage λ genome.</p

Clinical isolates and strains used in this study.

<p>(+) amplification of the 427-bp fragment; (-) no amplification.</p

Gene features of the IS<i>1245</i> flanking regions in pMA100.

<p>The similarities identified in the 7,525-bp sequenced fragment of pMA100 are indicated. Similarities at nucleotide levels were identified using BLASTn and the regions of similarity are depicted as hatched bars; identities at amino acid level were identified using BLASTx and each region is shown as a white arrow, indicating the direction of transcription. Black bar localizes the IS<i>1245</i> element. KpnI restriction sites are indicated.</p

Numbers of ORFs identified in <i>A. deanei</i> and <i>S. culicis</i> and their symbionts, according to the mechanisms of DNA replication and repair, signal transduction, transcription and translation.

<p>Numbers of ORFs identified in <i>A. deanei</i> and <i>S. culicis</i> and their symbionts, according to the mechanisms of DNA replication and repair, signal transduction, transcription and translation.</p

Summary of the origin of ORFs found in <i>A. deanei</i> and <i>S. culicis.</i>

*<p>Number of genes with identity to Prokaryotes.</p>**<p>Number of genes with identity to Eukaryotes.</p>***<p>Ratio of the number of genes with identity to Prokaryotes/Eukaryotes.</p

Genome alignments.

<p>The figure shows the alignment of the <i>A. deanei</i> endosymbiont (Endo-<i>A. deanei</i>) and the <i>S. culicis</i> endosymbiont (Endo-<i>S. culicis</i>) (A); between Endo-<i>A. deanei</i> and <i>T. asinigenitalis</i> (B), <i>T. equigenitalis</i> (C), or <i>Wolbachia</i> (D); and between <i>Wolbachia</i> and <i>T. asinigenitalis</i> (E). Alignments were performed with the ACT program based on tblastx analyses. Red (direct similarity) and blue lines (indirect similarity) connect similar regions with at least 700 bp and a score cutoff of 700. The numbers on the right indicate the size of the entire sequence for each organism.</p

Respiratory chain complexes identified in the predicted proteome of <i>A. deanei</i>, <i>S. culicis</i> and their respective endosymbionts.

*<p>The complex IV of the endosymbionts might be a cytochrome <i>d</i> ubiquinol oxidase identified in both organisms, instead a classical cytochrome <i>c</i> oxidase.</p