Plasmid Preparation:Article Title: Broadly Conserved Fungal Effector BEC1019 Suppresses Host Cell Death and Enhances Pathogen Virulence in Powdery Mildew of Barley (Hordeum vulgare L.) (Retracted)
Article Snippet: Bacterial strains and plasmids used (Whigham et al. MPMI 2015) Strain or plasmid Relevant characteristicsa,b Reference or source E. coli DB3.1 F– gyrA462 endA1 Δ(sr1-recA) mcrB mrr hsdS20(rB–, mB–) supE44 ara-14 galK2 lacY1 proA2 rpsL20(SmR) xyl-5 λ– leu mtl1 Invitrogen, Carlsbad, CA, USA TOP10 F- mcrA Δ (mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(araleu) 7697 galU galK rpsL (SmR) endA1 nupG Invitrogen, Carlsbad, CA, USA Xanthomonas campestris pv. raphani 756C CmR derivate of strain 756 Kamoun and Kado, 1990 HW9 T3S– derivative of 756C; carries insertion in hrpX; ApR, CmR, KmR Kamoun et al., 1992 Xanthomonas oryzae pv. oryzicola BLS256 Wild type; Philippines isolate Bogdanove et al., 2011 Plasmids pYM5 Gateway destination vector with pDD62 backbone; carries lac promoter followed by coding sequence for AvrBs2 T3S signal region (aa 1-97); contains ccdB gene; CmR, KmR This study pYMAJ1 pYM5 with the Gateway cassette replaced by a short insert that introduces an in frame stop codon immediatley following the coding sequence for aa 1-97 of AvrBs2, and a diagnostic EcoRI site; KmR This study pCR8/GW/TOPO Gateway entry vector; SpR Invitrogen, Carlsbad, CA, USA pYM5-BEC1019 pYM5 containing coding sequence for all but the first 18 residues (the predicted signal peptide) of BEC1019; KmR This study pVSP61-AvrPphB KmR Simonich and Innes, 1995 pYM5-Pra1 pYM5 containing coding sequence for Pra1 without its predicted signal peptide; KmR This study pYM5-BEC1019:E148A pYM5-BEC1019 with the E148A mutation; KmR This study pYM5-BEC1019:T149A pYM5-BEC1019 with the T149A mutation; KmR This study pYM5-BEC1019:V150A pYM5-BEC1019 with the V150A mutation; KmR This study pYM5-BEC1019:I151A pYM5-BEC1019 with the I151A mutation; KmR This study pYM5-BEC1019:C152S pYM5-BEC1019 with the C152S mutation; KmR This study pYM5-BEC1019:HRH-AAA pYM5-BEC1019 with the triple mutation H187A, R188A, and H191A; KmR This study a ApR, ampicillin resistant; CmR, chloramphenicol resistant; KmR, kanamycin resistant; SmR, streptomycin resistant; SpR, spectinomycin resistant b For mutations in BEC1019, single letter codes for resulting amino acids are used.
Sequencing:Article Title: Broadly Conserved Fungal Effector BEC1019 Suppresses Host Cell Death and Enhances Pathogen Virulence in Powdery Mildew of Barley (Hordeum vulgare L.) (Retracted)
Article Snippet: Bacterial strains and plasmids used (Whigham et al. MPMI 2015) Strain or plasmid Relevant characteristicsa,b Reference or source E. coli DB3.1 F– gyrA462 endA1 Δ(sr1-recA) mcrB mrr hsdS20(rB–, mB–) supE44 ara-14 galK2 lacY1 proA2 rpsL20(SmR) xyl-5 λ– leu mtl1 Invitrogen, Carlsbad, CA, USA TOP10 F- mcrA Δ (mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(araleu) 7697 galU galK rpsL (SmR) endA1 nupG Invitrogen, Carlsbad, CA, USA Xanthomonas campestris pv. raphani 756C CmR derivate of strain 756 Kamoun and Kado, 1990 HW9 T3S– derivative of 756C; carries insertion in hrpX; ApR, CmR, KmR Kamoun et al., 1992 Xanthomonas oryzae pv. oryzicola BLS256 Wild type; Philippines isolate Bogdanove et al., 2011 Plasmids pYM5 Gateway destination vector with pDD62 backbone; carries lac promoter followed by coding sequence for AvrBs2 T3S signal region (aa 1-97); contains ccdB gene; CmR, KmR This study pYMAJ1 pYM5 with the Gateway cassette replaced by a short insert that introduces an in frame stop codon immediatley following the coding sequence for aa 1-97 of AvrBs2, and a diagnostic EcoRI site; KmR This study pCR8/GW/TOPO Gateway entry vector; SpR Invitrogen, Carlsbad, CA, USA pYM5-BEC1019 pYM5 containing coding sequence for all but the first 18 residues (the predicted signal peptide) of BEC1019; KmR This study pVSP61-AvrPphB KmR Simonich and Innes, 1995 pYM5-Pra1 pYM5 containing coding sequence for Pra1 without its predicted signal peptide; KmR This study pYM5-BEC1019:E148A pYM5-BEC1019 with the E148A mutation; KmR This study pYM5-BEC1019:T149A pYM5-BEC1019 with the T149A mutation; KmR This study pYM5-BEC1019:V150A pYM5-BEC1019 with the V150A mutation; KmR This study pYM5-BEC1019:I151A pYM5-BEC1019 with the I151A mutation; KmR This study pYM5-BEC1019:C152S pYM5-BEC1019 with the C152S mutation; KmR This study pYM5-BEC1019:HRH-AAA pYM5-BEC1019 with the triple mutation H187A, R188A, and H191A; KmR This study a ApR, ampicillin resistant; CmR, chloramphenicol resistant; KmR, kanamycin resistant; SmR, streptomycin resistant; SpR, spectinomycin resistant b For mutations in BEC1019, single letter codes for resulting amino acids are used.
Diagnostic Assay:Article Title: Broadly Conserved Fungal Effector BEC1019 Suppresses Host Cell Death and Enhances Pathogen Virulence in Powdery Mildew of Barley (Hordeum vulgare L.) (Retracted)
Article Snippet: Bacterial strains and plasmids used (Whigham et al. MPMI 2015) Strain or plasmid Relevant characteristicsa,b Reference or source E. coli DB3.1 F– gyrA462 endA1 Δ(sr1-recA) mcrB mrr hsdS20(rB–, mB–) supE44 ara-14 galK2 lacY1 proA2 rpsL20(SmR) xyl-5 λ– leu mtl1 Invitrogen, Carlsbad, CA, USA TOP10 F- mcrA Δ (mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(araleu) 7697 galU galK rpsL (SmR) endA1 nupG Invitrogen, Carlsbad, CA, USA Xanthomonas campestris pv. raphani 756C CmR derivate of strain 756 Kamoun and Kado, 1990 HW9 T3S– derivative of 756C; carries insertion in hrpX; ApR, CmR, KmR Kamoun et al., 1992 Xanthomonas oryzae pv. oryzicola BLS256 Wild type; Philippines isolate Bogdanove et al., 2011 Plasmids pYM5 Gateway destination vector with pDD62 backbone; carries lac promoter followed by coding sequence for AvrBs2 T3S signal region (aa 1-97); contains ccdB gene; CmR, KmR This study pYMAJ1 pYM5 with the Gateway cassette replaced by a short insert that introduces an in frame stop codon immediatley following the coding sequence for aa 1-97 of AvrBs2, and a diagnostic EcoRI site; KmR This study pCR8/GW/TOPO Gateway entry vector; SpR Invitrogen, Carlsbad, CA, USA pYM5-BEC1019 pYM5 containing coding sequence for all but the first 18 residues (the predicted signal peptide) of BEC1019; KmR This study pVSP61-AvrPphB KmR Simonich and Innes, 1995 pYM5-Pra1 pYM5 containing coding sequence for Pra1 without its predicted signal peptide; KmR This study pYM5-BEC1019:E148A pYM5-BEC1019 with the E148A mutation; KmR This study pYM5-BEC1019:T149A pYM5-BEC1019 with the T149A mutation; KmR This study pYM5-BEC1019:V150A pYM5-BEC1019 with the V150A mutation; KmR This study pYM5-BEC1019:I151A pYM5-BEC1019 with the I151A mutation; KmR This study pYM5-BEC1019:C152S pYM5-BEC1019 with the C152S mutation; KmR This study pYM5-BEC1019:HRH-AAA pYM5-BEC1019 with the triple mutation H187A, R188A, and H191A; KmR This study a ApR, ampicillin resistant; CmR, chloramphenicol resistant; KmR, kanamycin resistant; SmR, streptomycin resistant; SpR, spectinomycin resistant b For mutations in BEC1019, single letter codes for resulting amino acids are used.
Mutagenesis:Article Title: Broadly Conserved Fungal Effector BEC1019 Suppresses Host Cell Death and Enhances Pathogen Virulence in Powdery Mildew of Barley (Hordeum vulgare L.) (Retracted)
Article Snippet: Bacterial strains and plasmids used (Whigham et al. MPMI 2015) Strain or plasmid Relevant characteristicsa,b Reference or source E. coli DB3.1 F– gyrA462 endA1 Δ(sr1-recA) mcrB mrr hsdS20(rB–, mB–) supE44 ara-14 galK2 lacY1 proA2 rpsL20(SmR) xyl-5 λ– leu mtl1 Invitrogen, Carlsbad, CA, USA TOP10 F- mcrA Δ (mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(araleu) 7697 galU galK rpsL (SmR) endA1 nupG Invitrogen, Carlsbad, CA, USA Xanthomonas campestris pv. raphani 756C CmR derivate of strain 756 Kamoun and Kado, 1990 HW9 T3S– derivative of 756C; carries insertion in hrpX; ApR, CmR, KmR Kamoun et al., 1992 Xanthomonas oryzae pv. oryzicola BLS256 Wild type; Philippines isolate Bogdanove et al., 2011 Plasmids pYM5 Gateway destination vector with pDD62 backbone; carries lac promoter followed by coding sequence for AvrBs2 T3S signal region (aa 1-97); contains ccdB gene; CmR, KmR This study pYMAJ1 pYM5 with the Gateway cassette replaced by a short insert that introduces an in frame stop codon immediatley following the coding sequence for aa 1-97 of AvrBs2, and a diagnostic EcoRI site; KmR This study pCR8/GW/TOPO Gateway entry vector; SpR Invitrogen, Carlsbad, CA, USA pYM5-BEC1019 pYM5 containing coding sequence for all but the first 18 residues (the predicted signal peptide) of BEC1019; KmR This study pVSP61-AvrPphB KmR Simonich and Innes, 1995 pYM5-Pra1 pYM5 containing coding sequence for Pra1 without its predicted signal peptide; KmR This study pYM5-BEC1019:E148A pYM5-BEC1019 with the E148A mutation; KmR This study pYM5-BEC1019:T149A pYM5-BEC1019 with the T149A mutation; KmR This study pYM5-BEC1019:V150A pYM5-BEC1019 with the V150A mutation; KmR This study pYM5-BEC1019:I151A pYM5-BEC1019 with the I151A mutation; KmR This study pYM5-BEC1019:C152S pYM5-BEC1019 with the C152S mutation; KmR This study pYM5-BEC1019:HRH-AAA pYM5-BEC1019 with the triple mutation H187A, R188A, and H191A; KmR This study a ApR, ampicillin resistant; CmR, chloramphenicol resistant; KmR, kanamycin resistant; SmR, streptomycin resistant; SpR, spectinomycin resistant b For mutations in BEC1019, single letter codes for resulting amino acids are used.
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