pgem-t , cloning vector with t overhangs, lacz ap r  (Promega)

Journal: Journal of Bacteriology

Article Title: Sulfide Consumption in Sulfurimonas denitrificans and Heterologous Expression of Its Three Sulfide-Quinone Reductase Homologs

doi: 10.1128/JB.01021-15

Figure Lengend Snippet: Strains and plasmids used in this study

Article Snippet: pGEM-T , Cloning vector with T overhangs, lacZ Ap r , Promega.

Techniques: Plasmid Preparation, Conjugation Assay, Clone Assay

Journal: PLoS ONE

Article Title: Diguanylate Cyclase Null Mutant Reveals That C-Di-GMP Pathway Regulates the Motility and Adherence of the Extremophile Bacterium Acidithiobacillus caldus

doi: 10.1371/journal.pone.0116399

Figure Lengend Snippet: Bacterial strains, plasmids and primers used in this study.

Article Snippet: pGEM-T , Ap r ; T-tailed PCR product cloning vector , Promega.

Techniques: Expressing, Plasmid Preparation, Clone Assay

Journal: Journal of Bacteriology

Article Title: Involvement of Regulatory Interactions among Global Regulators GlxR, SugR, and RamA in Expression of ramA in Corynebacterium glutamicum

doi: 10.1128/JB.00016-13

Figure Lengend Snippet: Bacterial strains and plasmids used in this study

Article Snippet: pGEM-T Easy , Ap r ; TA cloning vector , Promega.

Techniques: Plasmid Preparation, Binding Assay, Expressing, TA Cloning

Journal:

Article Title: Apyrase, the Product of the Virulence Plasmid-Encoded phoN2 ( apy ) Gene of Shigella flexneri , Is Necessary for Proper Unipolar IcsA Localization and for Efficient Intercellular Spread †

doi: 10.1128/JB.188.4.1620-1627.2006

Figure Lengend Snippet: Bacterial strains and plasmids a

Article Snippet: The locations of 3× FLAG-tagged OspB recombinant protein as well as of IpaB, IpaC, and apyrase proteins are indicated. na, not added. table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Strain or plasmid Relevant genotype or characteristic(s) Source or reference Strains M90T Wild-type S. flexneri serotype 5a 32 HND115 M90T Δ phoN2 ; susceptible This work HND201 M90T Δ ospB ; susceptible This work HND215 M90T Δ ospBphoN2 ; susceptible This work HND43 M90T Δ mxiA ; susceptible This work HND53 M90T Δ virB :: aphA-3 ; Km r This work HND549 M90T ospB -3×FLAG; susceptible This work HND5311 M90T Δ mxiA ospB -3×FLAG; susceptible This work SC560 M90T Δ icsA ::Ω; Sm r 10 M15[pREP4] E. coli K-12 strain carrying plasmid pREP4 Qiagen Plasmids pGEM-T Cloning vector, Ap r Promega pBluescript SK Cloning vector, Ap r Stratagene Inc., La Jolla, CA pBAD28 Arabinose-inducible P BAD expression vector, Ap r Cm r 13 pSUB11 Template plasmid carrying a 3×FLAG epitope and Km r cassette 42 pKD46 λ Red helper plasmid; oriR101 repA101 (Ts) P-araB-gam-bet-exo Ap r 7 pKD4 Template plasmid carrying a Km r gene with FLP recognition target sequence 7 pCP20 FLP helper plasmid; pSC101 replicon (Ts) bla cat Flp (λR p ) cI 857 Ap r Cm r 7 pREP4 lacI , Km r Qiagen pQE30 Cloning vector; Ap r Qiagen pHN301 pQE30 carrying His6- phoN2 ; Ap r This work pHN28 pBAD28 carrying phoN2 ; Ap r Cm r This work pHN111 pBAD28 carrying mxiA ; Ap r Cm r This work pFB1-4 pBluescript SK carrying a mutated phoN2 gene of EIEC strain HN280 presenting the R192P amino acid substitution 36 Open in a separate window a Ap r , ampicillin resistance; Cm r , cloramphenicol resistance; Km r , kanamycin resistance.

Techniques: Plasmid Preparation, Clone Assay, Expressing, Sequencing

Journal:

Article Title: Apyrase, the Product of the Virulence Plasmid-Encoded phoN2 ( apy ) Gene of Shigella flexneri , Is Necessary for Proper Unipolar IcsA Localization and for Efficient Intercellular Spread †

doi: 10.1128/JB.188.4.1620-1627.2006

Figure Lengend Snippet: Formation of plaques on confluent monolayers of HeLa and Caco-2 cells a

Article Snippet: The locations of 3× FLAG-tagged OspB recombinant protein as well as of IpaB, IpaC, and apyrase proteins are indicated. na, not added. table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Strain or plasmid Relevant genotype or characteristic(s) Source or reference Strains M90T Wild-type S. flexneri serotype 5a 32 HND115 M90T Δ phoN2 ; susceptible This work HND201 M90T Δ ospB ; susceptible This work HND215 M90T Δ ospBphoN2 ; susceptible This work HND43 M90T Δ mxiA ; susceptible This work HND53 M90T Δ virB :: aphA-3 ; Km r This work HND549 M90T ospB -3×FLAG; susceptible This work HND5311 M90T Δ mxiA ospB -3×FLAG; susceptible This work SC560 M90T Δ icsA ::Ω; Sm r 10 M15[pREP4] E. coli K-12 strain carrying plasmid pREP4 Qiagen Plasmids pGEM-T Cloning vector, Ap r Promega pBluescript SK Cloning vector, Ap r Stratagene Inc., La Jolla, CA pBAD28 Arabinose-inducible P BAD expression vector, Ap r Cm r 13 pSUB11 Template plasmid carrying a 3×FLAG epitope and Km r cassette 42 pKD46 λ Red helper plasmid; oriR101 repA101 (Ts) P-araB-gam-bet-exo Ap r 7 pKD4 Template plasmid carrying a Km r gene with FLP recognition target sequence 7 pCP20 FLP helper plasmid; pSC101 replicon (Ts) bla cat Flp (λR p ) cI 857 Ap r Cm r 7 pREP4 lacI , Km r Qiagen pQE30 Cloning vector; Ap r Qiagen pHN301 pQE30 carrying His6- phoN2 ; Ap r This work pHN28 pBAD28 carrying phoN2 ; Ap r Cm r This work pHN111 pBAD28 carrying mxiA ; Ap r Cm r This work pFB1-4 pBluescript SK carrying a mutated phoN2 gene of EIEC strain HN280 presenting the R192P amino acid substitution 36 Open in a separate window a Ap r , ampicillin resistance; Cm r , cloramphenicol resistance; Km r , kanamycin resistance.

Techniques:

Journal:

Article Title: Apyrase, the Product of the Virulence Plasmid-Encoded phoN2 ( apy ) Gene of Shigella flexneri , Is Necessary for Proper Unipolar IcsA Localization and for Efficient Intercellular Spread †

doi: 10.1128/JB.188.4.1620-1627.2006

Figure Lengend Snippet: phoN2 mutant strain HND115 forms plaques of approximately wild-type size on Caco-2 cell monolayers when complemented with plasmid pFB1-4. pFB1-4 (Table ​(Table1)1) contains a randomly generated mutant of the apy gene, the orthologue of phoN2 in the EIEC strain HN280, encoding a recombinant apyrase presenting the R192P substitution which inactivates its dNTP-hydrolyzing activity. (A) Western blot analysis with anti-apyrase antibodies of whole-cell extracts of M90T, HND115 (phoN2), and HND115/pFB1-4 (pBluescript SKapyR192P). (B) Plaques formed by M90T, HND115 (phoN2), and HND115/pFB1-4 (pBluescript SKapyR192P). Dishes were photographed after 48 h of infection.

Article Snippet: The locations of 3× FLAG-tagged OspB recombinant protein as well as of IpaB, IpaC, and apyrase proteins are indicated. na, not added. table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Strain or plasmid Relevant genotype or characteristic(s) Source or reference Strains M90T Wild-type S. flexneri serotype 5a 32 HND115 M90T Δ phoN2 ; susceptible This work HND201 M90T Δ ospB ; susceptible This work HND215 M90T Δ ospBphoN2 ; susceptible This work HND43 M90T Δ mxiA ; susceptible This work HND53 M90T Δ virB :: aphA-3 ; Km r This work HND549 M90T ospB -3×FLAG; susceptible This work HND5311 M90T Δ mxiA ospB -3×FLAG; susceptible This work SC560 M90T Δ icsA ::Ω; Sm r 10 M15[pREP4] E. coli K-12 strain carrying plasmid pREP4 Qiagen Plasmids pGEM-T Cloning vector, Ap r Promega pBluescript SK Cloning vector, Ap r Stratagene Inc., La Jolla, CA pBAD28 Arabinose-inducible P BAD expression vector, Ap r Cm r 13 pSUB11 Template plasmid carrying a 3×FLAG epitope and Km r cassette 42 pKD46 λ Red helper plasmid; oriR101 repA101 (Ts) P-araB-gam-bet-exo Ap r 7 pKD4 Template plasmid carrying a Km r gene with FLP recognition target sequence 7 pCP20 FLP helper plasmid; pSC101 replicon (Ts) bla cat Flp (λR p ) cI 857 Ap r Cm r 7 pREP4 lacI , Km r Qiagen pQE30 Cloning vector; Ap r Qiagen pHN301 pQE30 carrying His6- phoN2 ; Ap r This work pHN28 pBAD28 carrying phoN2 ; Ap r Cm r This work pHN111 pBAD28 carrying mxiA ; Ap r Cm r This work pFB1-4 pBluescript SK carrying a mutated phoN2 gene of EIEC strain HN280 presenting the R192P amino acid substitution 36 Open in a separate window a Ap r , ampicillin resistance; Cm r , cloramphenicol resistance; Km r , kanamycin resistance.

Techniques: Mutagenesis, Plasmid Preparation, Generated, Recombinant, Activity Assay, Western Blot, Infection

Journal:

Article Title: Novel Insertion Sequence Elements Associated with Genetic Heterogeneity and Phenotype Conversion in Ralstonia solanacearum

doi:

Figure Lengend Snippet: Bacterial strains and plasmids

Article Snippet: In this study, ACH0158-M81C (Table ) was identified as a PC mutant resulting from the insertion of IS Rso4 within phcA ( 1 , 2 ). table ft1 table-wrap mode="anchored" t5 caption a7 R. solanacearum strain or plasmid Relevant characteristic(s) a Source or reference b Strains ACH0158 Wild-type strain isolated from potato A. C. Hayward ACH0158-M81C Spontaneous PC mutant of ACH0158 This study ACH0158-M3 Spontaneous PC mutant of ACH0158 This study ACH0158-M8 Spontaneous PC mutant of ACH0158 This study ACH1061 Wild-type strain isolated from potato A. C. Hayward ACH1068S Wild-type strain isolated from potato A. C. Hayward CIP418 Wild-type strain isolated from peanut CIP CIP419 Wild-type strain isolated from peanut CIP Plasmids pKS(+), pKS(−), pSK(+), pSK(−) Bluescript vectors; Ap r Stratagene pUC19 Cloning vector; Ap r New England Biolabs pGEM-T Easy Cloning vector; Ap r Promega pISBE 5.2-kb Eco RI fragment containing IS Rso2 from wild-type ACH0158 cloned into pKS; Ap r This study pSV102 2.7-kb Eco RI fragment containing IS Rso4 and IS Rso3 from wild-type ACH0158 cloned pKS; Ap r This study pWT 1.46-kb Eco RI fragment containing phcA from wild-type ACH0158 cloned into pGEM-T Easy; Ap r This study pM81C 1.64-kb Eco RI fragment containing phcA81C from PC mutant ACH0158-M81C cloned into pGEM-T Easy; Ap r This study pM3 1.33-kb Eco RI fragment containing phcA3 from PC mutant ACH0158-M3 cloned into pGEM-T Easy; Ap r This study pM8 1.46-kb Eco RI fragment containing phcA8 from PC mutant ACH0158-M8 cloned into pGEM-T Easy; Ap r This study Open in a separate window a Ap r , resistance to ampicillin. b CIP, International Potato Centre, Lima, Peru.

Techniques: Plasmid Preparation, Isolation, Mutagenesis, Cloning, Clone Assay

Journal:

Article Title: Identification and Characterization of Two Chemotactic Transducers for Inorganic Phosphate in Pseudomonas aeruginosa

doi:

Figure Lengend Snippet: Bacterial strains and plasmids used in this study

Article Snippet: For P i limitation, P. aeruginosa cells grown overnight in 2× YT medium were inoculated (a 2.5% inoculum) into T 0 medium, which was prepared by omitting P i from T 5 medium, and incubated at 37°C with shaking. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Strain or plasmid Relevant characteristics Reference or source E. coli MV1184 ara Δ( lac-proAB ) rpsL thi (φ80 lacZ ΔM15) Δ( srl-recA ) 306 ::Tn 10 (Tc r ) F′[ traD36 proAB + lacl q lacZ ΔM15] 23 HB101 hsdS20 recA13 proA2 leu6 thi-1 rpsL20 ara-14 galK2 lacY1 xyl-5 mtl-1 supE44 3 P. aeruginosa PAO1 Prototroph, FP − (sex factor minus) 8 PP1 PAO1 derivative, ctpH :: kan This study PP2 PAO1 derivative, ctpL :: kan This study PP3 PAO1 derivative, ctpH :: tet ctpL :: kan This study APC1 PAO1 derivative, phoU 12 PHOB1 PAO1 derivative, phoB :: kan 12 PP4 PAO1 derivative, phoU ctpH :: kan This study PP5 PAO1 derivative, phoU ctpL :: kan This study PP6 PAO1 derivative, phoB :: kan ctpH :: tet This study PP7 PAO1 derivative, phoB :: kan ctpL :: tet This study Plasmids pGEM-T Easy PCR cloning vector; Ap r lacPOZ ′ Promega pUC118Tc pUC118 containing a 1.3-kb tet cartridge from pBR322; Ap r Tc r This study pUC4K pUC4 containing a 1.3-kb kan cartridge; Ap r Km r Pharmacia pCP19 Broad-host-range cosmid; Tc r IncP 6 pPT10.1 pGEM-T Easy with a 3.6-kb PCR fragment containing ctpH ; Ap r This study pPT11.1 pGEM-T Easy with a 3.3-kb PCR fragment containing ctpL ; Ap r This study pPT10.2 pPT10.1 derivative containing a 1.3-kb kan cartridge This study pPT11.2 pPT11.1 derivative containing a 1.3-kb kan cartridge This study pPT10.3 pPT10.1 derivative containing a 1.3-kb tet cartridge This study pPT10.4 pCP19 derivative containing a 3.6-kb Eco RI fragment of pPT10.1; Tc r This study pPT11.4 pCP19 derivative containing a 2.3-kb Eco RV fragment of pPT11.1; Tc r This study pKZ27 Broad-host-range transcriptional fusion vector; Km r lacZ IncQ This study pKZ27.1 pKZ27 containing a 1.2-kb fragment with amp gene from pBR322; Km r Cb r This study pPT10.5 pKZ27 derivative containing a 1.8-kb Kpn I fragment of pPT10.1; Km r This study pPT10.6 pKZ27.1 derivative containing a 1.8-kb Kpn I fragment of pPT10.1; Km r Cb r This study pPT11.5 pKZ27 derivative containing a 1.6-kb Eco RV- Sal I fragment of pPT10.1; Km r This study pPT11.6 pKZ27.1 derivative containing a 1.6-kb Eco RV- Sal I fragment of pPT10.1; Km r Cb r This study Open in a separate window Bacterial strains and plasmids used in this study

Techniques: Plasmid Preparation, PCR Cloning

Journal:

Article Title: Molecular and Functional Characterization of the Rhodopseudomonas palustris No. 7 Phosphoenolpyruvate Carboxykinase Gene

doi:

Figure Lengend Snippet: Bacterial strains and plasmids used in this study

Article Snippet: pGEM-T , Ap r ; α-lac/MCS; PCR cloning vector , Promega.

Techniques: Plasmid Preparation, Mutagenesis, PCR Cloning, Clone Assay