caption a7 strain plasmid intermediate relevant characteristic reference  (ATCC)

Journal: Infection and Immunity

Article Title: The Intermediate Region of Helicobacter pylori VacA Is a Determinant of Toxin Potency in a Jurkat T Cell Assay

doi: 10.1128/IAI.00052-12

Figure Lengend Snippet: Amino acid sequences of the VacA i-region in different H. pylori strains. The secreted p88 VacA protein (comprising 821 amino acids in H. pylori strain 60190) contains two domains, designated p33 and p55. Amino acid sequence variation in VacA proteins from different H. pylori strains is especially prominent in three regions, which are designated s-, i-, and m-regions. The i-region and a portion of the s-region are localized within the p33 domain, and the m-region is localized within the p55 domain. The lower portion of this figure shows the sequences of VacA i-regions from strain 60190 (GenBank no. U05676), Tx30a (GenBank no. U29401), and X47. The figure illustrates the sequence of the secreted VacA protein of H. pylori strain 60190 (amino acids G120 to L236) and corresponding regions of VacA in two other wild-type strains (Tx30a and X47). Clusters of amino acid polymorphisms defined as clusters A, B, and C are indicated. Asterisks (*) indicate amino acids that are identical in all three sequences. The sequence of VacA from strain 60190 is designated a prototype for the i1 region, and the sequence of VacA from strain Tx30a is designated a prototype for the i2 region. VacA from strain X47 contains a type i2 sequence in cluster C, and cluster B is chimeric.

Article Snippet: H. pylori liquid cultures were grown in brucella broth supplemented with either activated charcoal or 5% fetal bovine serum (FBS) ( 11 ). table ft1 table-wrap mode="anchored" t5 caption a7 Strain/plasmid Relevant characteristics Reference Strains 60190 Wild type (ATCC 49503); vacA s1/i1/m1 40 60190 Δ rdxA Same as 60190 except HP0954 ( rdxA ) gene deleted; metronidazole resistant This study 60190 vacA :: cat rdxA Same as 60190 Δ rdxA except cat cassette and rdxA inserted in vacA ; chloramphenicol resistant and metronidazole sensitive; expression of VacA is disrupted This study 60190 i2B Same as 60190 Δ rdxA except vacA cluster B changed to i2 This study 60190 i2C Same as 60190 Δ rdxA except vacA cluster C changed to i2 This study 60190 i2BC Same as 60190 Δ rdxA except vacA clusters B and C changed to i2 This study 60190 i1/i2C Same as 60190 Δ rdxA except vacA cluster C has 4 amino acids changed to i2 This study X47 Wild type; vacA s1/m2, chimeric i-region 32 X47 Δ rdxA Same as X47 except HP0954 ( rdxA ) gene deleted; metronidazole resistant This study X47 vacA :: cat rdxA Same as X47 Δ rdxA except cat cassette and rdxA inserted in vacA ; chloramphenicol resistant and metronidazole sensitive; expression of VacA is disrupted This study X47 i1C Same as X47 Δ rdxA except vacA cluster C changed to i1 This study Plasmids pMM672 Allows deletion of rdxA in H. pylori strains 41 pCGR1 Contains cat-rdxA cassette in StuI site; derived from pA178 plasmid This study pCGR2 Contains cat-rdxA cassette in EcoRV site from X47 vacA This study pCGR3 60910 cluster B changed from i1 to i2 by inverse PCR using primers B1F and B1R This study pCGR4 60190 cluster C changed from i1 to i2 by inverse PCR using primers C1F and C1R This study pCGR5 60190 clusters B and C changed from i1 to i2 by inverse PCR using primers C1F and C1R and pCGR3 as the template This study pCGR6 A portion of 60190 cluster C changed from i1 to i2 by inverse PCR using primers C2F and C2R This study pCGR7 X47 cluster C changed from i2 to i1 by inverse PCR using primers C3F and C3R This study p55 Expresses VacA p55 27 p33 Expresses VacA p33 31 p33 i2 Expresses p33 i2 This study Open in a separate window H. pylori strains and plasmids Preparation of H. pylori broth culture supernatants and normalization of VacA concentrations.

Techniques: Sequencing

Journal: Infection and Immunity

Article Title: The Intermediate Region of Helicobacter pylori VacA Is a Determinant of Toxin Potency in a Jurkat T Cell Assay

doi: 10.1128/IAI.00052-12

Figure Lengend Snippet: H. pylori strains and plasmids

Article Snippet: H. pylori liquid cultures were grown in brucella broth supplemented with either activated charcoal or 5% fetal bovine serum (FBS) ( 11 ). table ft1 table-wrap mode="anchored" t5 caption a7 Strain/plasmid Relevant characteristics Reference Strains 60190 Wild type (ATCC 49503); vacA s1/i1/m1 40 60190 Δ rdxA Same as 60190 except HP0954 ( rdxA ) gene deleted; metronidazole resistant This study 60190 vacA :: cat rdxA Same as 60190 Δ rdxA except cat cassette and rdxA inserted in vacA ; chloramphenicol resistant and metronidazole sensitive; expression of VacA is disrupted This study 60190 i2B Same as 60190 Δ rdxA except vacA cluster B changed to i2 This study 60190 i2C Same as 60190 Δ rdxA except vacA cluster C changed to i2 This study 60190 i2BC Same as 60190 Δ rdxA except vacA clusters B and C changed to i2 This study 60190 i1/i2C Same as 60190 Δ rdxA except vacA cluster C has 4 amino acids changed to i2 This study X47 Wild type; vacA s1/m2, chimeric i-region 32 X47 Δ rdxA Same as X47 except HP0954 ( rdxA ) gene deleted; metronidazole resistant This study X47 vacA :: cat rdxA Same as X47 Δ rdxA except cat cassette and rdxA inserted in vacA ; chloramphenicol resistant and metronidazole sensitive; expression of VacA is disrupted This study X47 i1C Same as X47 Δ rdxA except vacA cluster C changed to i1 This study Plasmids pMM672 Allows deletion of rdxA in H. pylori strains 41 pCGR1 Contains cat-rdxA cassette in StuI site; derived from pA178 plasmid This study pCGR2 Contains cat-rdxA cassette in EcoRV site from X47 vacA This study pCGR3 60910 cluster B changed from i1 to i2 by inverse PCR using primers B1F and B1R This study pCGR4 60190 cluster C changed from i1 to i2 by inverse PCR using primers C1F and C1R This study pCGR5 60190 clusters B and C changed from i1 to i2 by inverse PCR using primers C1F and C1R and pCGR3 as the template This study pCGR6 A portion of 60190 cluster C changed from i1 to i2 by inverse PCR using primers C2F and C2R This study pCGR7 X47 cluster C changed from i2 to i1 by inverse PCR using primers C3F and C3R This study p55 Expresses VacA p55 27 p33 Expresses VacA p33 31 p33 i2 Expresses p33 i2 This study Open in a separate window H. pylori strains and plasmids Preparation of H. pylori broth culture supernatants and normalization of VacA concentrations.

Techniques: Expressing, Derivative Assay, Plasmid Preparation, Inverse PCR

Journal:

Article Title: Aggregation Substance Increases Adherence and Internalization, but Not Translocation, of Enterococcus faecalis through Different Intestinal Epithelial Cells In Vitro

doi:

Figure Lengend Snippet: E. faecalis strains

Article Snippet: Appropriate dilutions of the suspensions were plated on nutrient agar in order to determine the CFU per milliliter of infection medium. table ft1 table-wrap mode="anchored" t5 caption a7 Strain Relevant characteristic Reference OG1X Plasmid free 13 OG1X/pAM721 pAD1 derivative containing Tn 917 insertion within regulatory region of asa-1 resulting in constitutive expression of AS 12 OG1X/pAM944 pAD1 derivative containing Tn 917 insertion in asa-1 resulting in secretion of truncated AS 7, 13 Open in a separate window E. faecalis strains Intestinal epithelial cell lines originating from the colon (HT 29 and T84), ileum (HCT-8), and duodenum (Hutu 80) were used at passages below 10 from a stock culture purchased from the American Type Culture Collection (Manassas, Va.).

Techniques: Plasmid Preparation, Expressing