B. Bakhshi, M.R. Pourshafie, F. Navabakbar, A. Tavakoli

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Genomic organisation of the CTX element among toxigenic Vibrio cholerae isolates B. Bakhshi, M.R. Pourshafie, F. Navabakbar, A. Tavakoli Clinical Microbiology and Infection Volume 14, Issue 6, Pages 562-568 (June 2008) DOI: 10.1111/j.1469-0691.2008.01976.x Copyright © 2008 European Society of Clinical Infectious Diseases Terms and Conditions

Fig. 1 Schematic arrangement of the structure of the CTX genetic element and the flanking regions of strain N16961. Open reading frames are shown as arrows. The black triangles represent CTXϕ attachment sites (attRS). Solid bars indicate intergenic regions flanking rstR; patterned bars indicate the attB1 and attB2 regions. Clinical Microbiology and Infection 2008 14, 562-568DOI: (10.1111/j.1469-0691.2008.01976.x) Copyright © 2008 European Society of Clinical Infectious Diseases Terms and Conditions

Fig. 2 Representative amplified DNA products obtained by long-PCR (L-PCR) using ig1-F/attB2-R primers. (a) Lanes 3, 2 and 1 are L-PCR products of patterns 3, 2 and 1, respectively. (b) Lanes 1, 2 and 3 show EcoRV digests of the L-PCR products of the three patterns. (c) Lanes 1 and 2 show DraI digests of the L-PCR products of patterns 1 and 2, respectively. (d) Lanes 1 and 2 show BglI digests and lanes 3 and 4 show XbaI digests of the L-PCR products of patterns 1 and 2. M, molecular size markers. Clinical Microbiology and Infection 2008 14, 562-568DOI: (10.1111/j.1469-0691.2008.01976.x) Copyright © 2008 European Society of Clinical Infectious Diseases Terms and Conditions

Fig. 3 Southern blot hybridisation patterns of the Vibrio cholerae isolates obtained using (a) ctx and (b) zot gene probes. Lanes: 1 and 2, profile 1; 3 and 4, profile 2; 5 and 6, profile 3. Clinical Microbiology and Infection 2008 14, 562-568DOI: (10.1111/j.1469-0691.2008.01976.x) Copyright © 2008 European Society of Clinical Infectious Diseases Terms and Conditions

Fig. 4 Proposed schematic genomic organisation of the CTX elements. The three possible arrangements found in all Vibrio cholerae isolates are labelled as 1, 2 and 3. The vertical arrows indicate the site of digestion by PstI. PstI restriction sites were determined from the fully sequenced V. cholerae strain N16961 [7] using NEB cutter software. , complete CTX genetic element; , TLC element; , RTX cluster; , truncated CTX genetic element; , RS1; , ctx probe; , zot probe. Clinical Microbiology and Infection 2008 14, 562-568DOI: (10.1111/j.1469-0691.2008.01976.x) Copyright © 2008 European Society of Clinical Infectious Diseases Terms and Conditions

Fig. 5 Pulsed-field gel electrophoresis patterns of the isolates obtained following digestion with NotI. M, molecular size marker. All Vibrio cholerae isolates belonged to either pulsotype 1 (lane 1) or pulsotype 2 (lane 2). Pulsotype 2 shows an additional 97-kb band. Clinical Microbiology and Infection 2008 14, 562-568DOI: (10.1111/j.1469-0691.2008.01976.x) Copyright © 2008 European Society of Clinical Infectious Diseases Terms and Conditions