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. 2000 Sep;68(9):4856-64.
doi: 10.1128/IAI.68.9.4856-4864.2000.

Structural analysis of phage-borne stx genes and their flanking sequences in shiga toxin-producing Escherichia coli and Shigella dysenteriae type 1 strains

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Structural analysis of phage-borne stx genes and their flanking sequences in shiga toxin-producing Escherichia coli and Shigella dysenteriae type 1 strains

A Unkmeir (V体育ios版) et al. Infect Immun. 2000 Sep.

Abstract

The stx-flanking regions of 49 Shiga toxin-producing Escherichia coli strains and nine Shigella dysenteriae serotype 1 strains containing either stx, stx(1), stx(2), or stx(2) variant genes, were examined. We analyzed these regions by PCR using a set of primers with one primer specific for the respective stx gene and a second primer complementary to sequences of Stx phages H-19B and 933W. We further characterized the amplification products by restriction endonuclease digestion and nucleotide sequencing. PCR products of stx(1)-containing E. coli strains of serogroups O157, O26, and 0103 showed the same lengths and similar restriction patterns. However, we failed to amplify the 3' stx-flanking region in stx(1)-harboring E. coli O111:H(-) strains. Stx2-producing E. coli strains revealed amplification products of different lengths and restriction patterns, suggesting greater heterogeneity than in stx(1)-positive strains. We also obtained specific PCR products for two Stx2c-producing and seven Stx2f-producing E. coli strains when they were subjected to PCR analysis. In nine S. dysenteriae type 1 strains, H-19B- and 933W-specific primers amplified only the 3' stx-flanking region VSports手机版. The results of our study demonstrate that the stx genes of all strains investigated are continuous with phage sequences. Whereas almost all strains except E. coli O111:H(-) strains were associated with a S-like gene, association with Q could not be demonstrated in nine S. dysenteriae type 1 strains and three E. coli strains. Furthermore, we showed that the organization of the stx-flanking regions is similar in all strains investigated, whereas fine-structure analysis showed subtle differences among the sequences examined. Our results support the hypothesis that stx genes in E. coli and S. dysenteriae are generally phage-borne. .

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Figures (VSports app下载)

FIG. 1
FIG. 1
Structure of the stx-flanking regions of Stx phages H-19B (A) and 933W (B) analyzed in this study. The arrows indicate the positions and directions of PCR and sequencing primers. The phage genes Roi, NinG, Q, stxA1, stxB1, stxA2, stxB2, and S are depicted by boxes. Lines indicate the expected PCR product length.
FIG. 2
FIG. 2
Structure of stx genes and stx-flanking sequences present in selected E. coli and S. dysenteriae strains. (A) Comparison of phage H-19B, E. coli O103:H2 strain 1858/96 and E. coli O111:H strain 1639/77. (B) Comparison of phage H-19B, S. dysenteriae type 1 strain H2765-34/81, and phage 933W. Arrows indicate the lengths and directions of open reading frames. The percentage of sequence identity of strain 1858/96 to phage H-19B is depicted as a lightly shaded box. The bar represents a length of 1 kb. Shaded bars below E. coli O111:H strain 1639/77 represent regions with sequence identities to phage H-19B, lambda, and S. enterica serovar Typhimurium. Gene designations are described in the text.
FIG. 3
FIG. 3
Structure of stx2 and stx2 variant genes and stx-flanking sequences present in selected E. coli strains. (A) Comparison of phage 933W with E. coli O26:H11 strain 1448/97, E. coli O26:H11 strain ED-147, and E. coli O145:H strain 3985/96. (B) Comparison of phage 933W with E. coli O157:H strain E32511, E. coli O145:H strain 4865/96, E. coli O128:B12 strain H.I.8, and E. coli O128:H2 T4/97. Symbols are the same as in Fig. 2.

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