Expressing:Article Title: The vesicle-forming host ESCRT complex promotes multiplication of
<i>Rhodococcus equi</i>
in macrophages
Article Snippet: .. 1Institute for Cell Biology, University of Bonn, Bonn, Germany 2Proteomics Core, Weill Cornell Medicine-Qatar, Doha, Qatar 3Institute for Medical Microbiology, Immunology and Hygiene, University of Cologne, Cologne, Germany 4Center for Molecular Medicine Cologne, University of Cologne, Cologne, Germany TABLE 4 Plasmids used in this study Gene/vector In vector Descriptiona Reference Mammalian CHMP3-YFP pEYFP-C1 Expressing Chmp3, acts dominant-negative because of EYFP fusion, Kanr Lambert et al. (84) CHMP4b-DsRed pDsRed-N1 Expressing Chmp4; acts dominant-negative because of DsRed fusion, Kanr Lambert et al. (84) DsRed pDsRed-N1 Expressing DsRed, Kanr Clontech EYFP pEYFP-C1 Expressing yellow fluorescent protein, Kanr Clontech GFP pEGFP-C1 Expressing enhanced green fluorescent protein, Kanr Clontech HGS/HGR-GFP pEGFP-C2 Expressing Chmp3; acts dominant-negative because of EGFP fusion, Kanr Tu et al. (85) VPS4A -GFP pEGFP-N1 Expressing a VPS4A-EGFP fusion protein, Kanr Tu et al.(85) VPS4A K173Q-GFP pEGFP-N1 Expressing a VPS4-EGFP fusion protein with point mutation in amino acid residue 173 of VPS4; acts as dominant-negative, Kanr Tu et al. (85) Bacterial vapA pET28a Expression vector for production of his6-tagged recombinant virulence-associated gene (VapA) from R. equi, lacking the secretion-mediating signal sequence, Kanr (8) aKanR, kanamycin resistant. .. order no. ALIX GUACCUCAGUCUAUACUUAtt s71368 CHMP3 GCUGGUUAAUGAAUGGUCAtt s83727 HRS/HGS AGUACAAGGUGGUCCAGGAtt s67517 TSG101 GCGUUAUCGAGGUAAUAUAtt s75514 VPS4A GAACAUACGGUGGAAUGAUtt s100252 VPS4B CGAAGAUUUGAGAAACGUAtt s73810 Mission siRNA universal negative control no. 1 = “scrambled siRNA” Sigma-Merck SIC001 Month XXXX Volume 0 Issue 0 10.1128/asmam.00010-2519 h ttp s: //j ou rn al s. as m .o rg /jo ur na l/a sm -a ni m al -m ic ro bi ol og y on 1 2 Fe br ua ry 2 02 6 by 2 40 0: 1a 00 :4 ba 5: 4c 42 :f 13 6: 57 9c :3 ae e: 68 f8
Dominant Negative Mutation:Article Title: The vesicle-forming host ESCRT complex promotes multiplication of
<i>Rhodococcus equi</i>
in macrophages
Article Snippet: .. 1Institute for Cell Biology, University of Bonn, Bonn, Germany 2Proteomics Core, Weill Cornell Medicine-Qatar, Doha, Qatar 3Institute for Medical Microbiology, Immunology and Hygiene, University of Cologne, Cologne, Germany 4Center for Molecular Medicine Cologne, University of Cologne, Cologne, Germany TABLE 4 Plasmids used in this study Gene/vector In vector Descriptiona Reference Mammalian CHMP3-YFP pEYFP-C1 Expressing Chmp3, acts dominant-negative because of EYFP fusion, Kanr Lambert et al. (84) CHMP4b-DsRed pDsRed-N1 Expressing Chmp4; acts dominant-negative because of DsRed fusion, Kanr Lambert et al. (84) DsRed pDsRed-N1 Expressing DsRed, Kanr Clontech EYFP pEYFP-C1 Expressing yellow fluorescent protein, Kanr Clontech GFP pEGFP-C1 Expressing enhanced green fluorescent protein, Kanr Clontech HGS/HGR-GFP pEGFP-C2 Expressing Chmp3; acts dominant-negative because of EGFP fusion, Kanr Tu et al. (85) VPS4A -GFP pEGFP-N1 Expressing a VPS4A-EGFP fusion protein, Kanr Tu et al.(85) VPS4A K173Q-GFP pEGFP-N1 Expressing a VPS4-EGFP fusion protein with point mutation in amino acid residue 173 of VPS4; acts as dominant-negative, Kanr Tu et al. (85) Bacterial vapA pET28a Expression vector for production of his6-tagged recombinant virulence-associated gene (VapA) from R. equi, lacking the secretion-mediating signal sequence, Kanr (8) aKanR, kanamycin resistant. .. order no. ALIX GUACCUCAGUCUAUACUUAtt s71368 CHMP3 GCUGGUUAAUGAAUGGUCAtt s83727 HRS/HGS AGUACAAGGUGGUCCAGGAtt s67517 TSG101 GCGUUAUCGAGGUAAUAUAtt s75514 VPS4A GAACAUACGGUGGAAUGAUtt s100252 VPS4B CGAAGAUUUGAGAAACGUAtt s73810 Mission siRNA universal negative control no. 1 = “scrambled siRNA” Sigma-Merck SIC001 Month XXXX Volume 0 Issue 0 10.1128/asmam.00010-2519 h ttp s: //j ou rn al s. as m .o rg /jo ur na l/a sm -a ni m al -m ic ro bi ol og y on 1 2 Fe br ua ry 2 02 6 by 2 40 0: 1a 00 :4 ba 5: 4c 42 :f 13 6: 57 9c :3 ae e: 68 f8
Mutagenesis:Article Title: The vesicle-forming host ESCRT complex promotes multiplication of
<i>Rhodococcus equi</i>
in macrophages
Article Snippet: .. 1Institute for Cell Biology, University of Bonn, Bonn, Germany 2Proteomics Core, Weill Cornell Medicine-Qatar, Doha, Qatar 3Institute for Medical Microbiology, Immunology and Hygiene, University of Cologne, Cologne, Germany 4Center for Molecular Medicine Cologne, University of Cologne, Cologne, Germany TABLE 4 Plasmids used in this study Gene/vector In vector Descriptiona Reference Mammalian CHMP3-YFP pEYFP-C1 Expressing Chmp3, acts dominant-negative because of EYFP fusion, Kanr Lambert et al. (84) CHMP4b-DsRed pDsRed-N1 Expressing Chmp4; acts dominant-negative because of DsRed fusion, Kanr Lambert et al. (84) DsRed pDsRed-N1 Expressing DsRed, Kanr Clontech EYFP pEYFP-C1 Expressing yellow fluorescent protein, Kanr Clontech GFP pEGFP-C1 Expressing enhanced green fluorescent protein, Kanr Clontech HGS/HGR-GFP pEGFP-C2 Expressing Chmp3; acts dominant-negative because of EGFP fusion, Kanr Tu et al. (85) VPS4A -GFP pEGFP-N1 Expressing a VPS4A-EGFP fusion protein, Kanr Tu et al.(85) VPS4A K173Q-GFP pEGFP-N1 Expressing a VPS4-EGFP fusion protein with point mutation in amino acid residue 173 of VPS4; acts as dominant-negative, Kanr Tu et al. (85) Bacterial vapA pET28a Expression vector for production of his6-tagged recombinant virulence-associated gene (VapA) from R. equi, lacking the secretion-mediating signal sequence, Kanr (8) aKanR, kanamycin resistant. .. order no. ALIX GUACCUCAGUCUAUACUUAtt s71368 CHMP3 GCUGGUUAAUGAAUGGUCAtt s83727 HRS/HGS AGUACAAGGUGGUCCAGGAtt s67517 TSG101 GCGUUAUCGAGGUAAUAUAtt s75514 VPS4A GAACAUACGGUGGAAUGAUtt s100252 VPS4B CGAAGAUUUGAGAAACGUAtt s73810 Mission siRNA universal negative control no. 1 = “scrambled siRNA” Sigma-Merck SIC001 Month XXXX Volume 0 Issue 0 10.1128/asmam.00010-2519 h ttp s: //j ou rn al s. as m .o rg /jo ur na l/a sm -a ni m al -m ic ro bi ol og y on 1 2 Fe br ua ry 2 02 6 by 2 40 0: 1a 00 :4 ba 5: 4c 42 :f 13 6: 57 9c :3 ae e: 68 f8
Residue:Article Title: The vesicle-forming host ESCRT complex promotes multiplication of
<i>Rhodococcus equi</i>
in macrophages
Article Snippet: .. 1Institute for Cell Biology, University of Bonn, Bonn, Germany 2Proteomics Core, Weill Cornell Medicine-Qatar, Doha, Qatar 3Institute for Medical Microbiology, Immunology and Hygiene, University of Cologne, Cologne, Germany 4Center for Molecular Medicine Cologne, University of Cologne, Cologne, Germany TABLE 4 Plasmids used in this study Gene/vector In vector Descriptiona Reference Mammalian CHMP3-YFP pEYFP-C1 Expressing Chmp3, acts dominant-negative because of EYFP fusion, Kanr Lambert et al. (84) CHMP4b-DsRed pDsRed-N1 Expressing Chmp4; acts dominant-negative because of DsRed fusion, Kanr Lambert et al. (84) DsRed pDsRed-N1 Expressing DsRed, Kanr Clontech EYFP pEYFP-C1 Expressing yellow fluorescent protein, Kanr Clontech GFP pEGFP-C1 Expressing enhanced green fluorescent protein, Kanr Clontech HGS/HGR-GFP pEGFP-C2 Expressing Chmp3; acts dominant-negative because of EGFP fusion, Kanr Tu et al. (85) VPS4A -GFP pEGFP-N1 Expressing a VPS4A-EGFP fusion protein, Kanr Tu et al.(85) VPS4A K173Q-GFP pEGFP-N1 Expressing a VPS4-EGFP fusion protein with point mutation in amino acid residue 173 of VPS4; acts as dominant-negative, Kanr Tu et al. (85) Bacterial vapA pET28a Expression vector for production of his6-tagged recombinant virulence-associated gene (VapA) from R. equi, lacking the secretion-mediating signal sequence, Kanr (8) aKanR, kanamycin resistant. .. order no. ALIX GUACCUCAGUCUAUACUUAtt s71368 CHMP3 GCUGGUUAAUGAAUGGUCAtt s83727 HRS/HGS AGUACAAGGUGGUCCAGGAtt s67517 TSG101 GCGUUAUCGAGGUAAUAUAtt s75514 VPS4A GAACAUACGGUGGAAUGAUtt s100252 VPS4B CGAAGAUUUGAGAAACGUAtt s73810 Mission siRNA universal negative control no. 1 = “scrambled siRNA” Sigma-Merck SIC001 Month XXXX Volume 0 Issue 0 10.1128/asmam.00010-2519 h ttp s: //j ou rn al s. as m .o rg /jo ur na l/a sm -a ni m al -m ic ro bi ol og y on 1 2 Fe br ua ry 2 02 6 by 2 40 0: 1a 00 :4 ba 5: 4c 42 :f 13 6: 57 9c :3 ae e: 68 f8
Plasmid Preparation:Article Title: The vesicle-forming host ESCRT complex promotes multiplication of
<i>Rhodococcus equi</i>
in macrophages
Article Snippet: .. 1Institute for Cell Biology, University of Bonn, Bonn, Germany 2Proteomics Core, Weill Cornell Medicine-Qatar, Doha, Qatar 3Institute for Medical Microbiology, Immunology and Hygiene, University of Cologne, Cologne, Germany 4Center for Molecular Medicine Cologne, University of Cologne, Cologne, Germany TABLE 4 Plasmids used in this study Gene/vector In vector Descriptiona Reference Mammalian CHMP3-YFP pEYFP-C1 Expressing Chmp3, acts dominant-negative because of EYFP fusion, Kanr Lambert et al. (84) CHMP4b-DsRed pDsRed-N1 Expressing Chmp4; acts dominant-negative because of DsRed fusion, Kanr Lambert et al. (84) DsRed pDsRed-N1 Expressing DsRed, Kanr Clontech EYFP pEYFP-C1 Expressing yellow fluorescent protein, Kanr Clontech GFP pEGFP-C1 Expressing enhanced green fluorescent protein, Kanr Clontech HGS/HGR-GFP pEGFP-C2 Expressing Chmp3; acts dominant-negative because of EGFP fusion, Kanr Tu et al. (85) VPS4A -GFP pEGFP-N1 Expressing a VPS4A-EGFP fusion protein, Kanr Tu et al.(85) VPS4A K173Q-GFP pEGFP-N1 Expressing a VPS4-EGFP fusion protein with point mutation in amino acid residue 173 of VPS4; acts as dominant-negative, Kanr Tu et al. (85) Bacterial vapA pET28a Expression vector for production of his6-tagged recombinant virulence-associated gene (VapA) from R. equi, lacking the secretion-mediating signal sequence, Kanr (8) aKanR, kanamycin resistant. .. order no. ALIX GUACCUCAGUCUAUACUUAtt s71368 CHMP3 GCUGGUUAAUGAAUGGUCAtt s83727 HRS/HGS AGUACAAGGUGGUCCAGGAtt s67517 TSG101 GCGUUAUCGAGGUAAUAUAtt s75514 VPS4A GAACAUACGGUGGAAUGAUtt s100252 VPS4B CGAAGAUUUGAGAAACGUAtt s73810 Mission siRNA universal negative control no. 1 = “scrambled siRNA” Sigma-Merck SIC001 Month XXXX Volume 0 Issue 0 10.1128/asmam.00010-2519 h ttp s: //j ou rn al s. as m .o rg /jo ur na l/a sm -a ni m al -m ic ro bi ol og y on 1 2 Fe br ua ry 2 02 6 by 2 40 0: 1a 00 :4 ba 5: 4c 42 :f 13 6: 57 9c :3 ae e: 68 f8
Recombinant:Article Title: The vesicle-forming host ESCRT complex promotes multiplication of
<i>Rhodococcus equi</i>
in macrophages
Article Snippet: .. 1Institute for Cell Biology, University of Bonn, Bonn, Germany 2Proteomics Core, Weill Cornell Medicine-Qatar, Doha, Qatar 3Institute for Medical Microbiology, Immunology and Hygiene, University of Cologne, Cologne, Germany 4Center for Molecular Medicine Cologne, University of Cologne, Cologne, Germany TABLE 4 Plasmids used in this study Gene/vector In vector Descriptiona Reference Mammalian CHMP3-YFP pEYFP-C1 Expressing Chmp3, acts dominant-negative because of EYFP fusion, Kanr Lambert et al. (84) CHMP4b-DsRed pDsRed-N1 Expressing Chmp4; acts dominant-negative because of DsRed fusion, Kanr Lambert et al. (84) DsRed pDsRed-N1 Expressing DsRed, Kanr Clontech EYFP pEYFP-C1 Expressing yellow fluorescent protein, Kanr Clontech GFP pEGFP-C1 Expressing enhanced green fluorescent protein, Kanr Clontech HGS/HGR-GFP pEGFP-C2 Expressing Chmp3; acts dominant-negative because of EGFP fusion, Kanr Tu et al. (85) VPS4A -GFP pEGFP-N1 Expressing a VPS4A-EGFP fusion protein, Kanr Tu et al.(85) VPS4A K173Q-GFP pEGFP-N1 Expressing a VPS4-EGFP fusion protein with point mutation in amino acid residue 173 of VPS4; acts as dominant-negative, Kanr Tu et al. (85) Bacterial vapA pET28a Expression vector for production of his6-tagged recombinant virulence-associated gene (VapA) from R. equi, lacking the secretion-mediating signal sequence, Kanr (8) aKanR, kanamycin resistant. .. order no. ALIX GUACCUCAGUCUAUACUUAtt s71368 CHMP3 GCUGGUUAAUGAAUGGUCAtt s83727 HRS/HGS AGUACAAGGUGGUCCAGGAtt s67517 TSG101 GCGUUAUCGAGGUAAUAUAtt s75514 VPS4A GAACAUACGGUGGAAUGAUtt s100252 VPS4B CGAAGAUUUGAGAAACGUAtt s73810 Mission siRNA universal negative control no. 1 = “scrambled siRNA” Sigma-Merck SIC001 Month XXXX Volume 0 Issue 0 10.1128/asmam.00010-2519 h ttp s: //j ou rn al s. as m .o rg /jo ur na l/a sm -a ni m al -m ic ro bi ol og y on 1 2 Fe br ua ry 2 02 6 by 2 40 0: 1a 00 :4 ba 5: 4c 42 :f 13 6: 57 9c :3 ae e: 68 f8
Sequencing:Article Title: The vesicle-forming host ESCRT complex promotes multiplication of
<i>Rhodococcus equi</i>
in macrophages
Article Snippet: .. 1Institute for Cell Biology, University of Bonn, Bonn, Germany 2Proteomics Core, Weill Cornell Medicine-Qatar, Doha, Qatar 3Institute for Medical Microbiology, Immunology and Hygiene, University of Cologne, Cologne, Germany 4Center for Molecular Medicine Cologne, University of Cologne, Cologne, Germany TABLE 4 Plasmids used in this study Gene/vector In vector Descriptiona Reference Mammalian CHMP3-YFP pEYFP-C1 Expressing Chmp3, acts dominant-negative because of EYFP fusion, Kanr Lambert et al. (84) CHMP4b-DsRed pDsRed-N1 Expressing Chmp4; acts dominant-negative because of DsRed fusion, Kanr Lambert et al. (84) DsRed pDsRed-N1 Expressing DsRed, Kanr Clontech EYFP pEYFP-C1 Expressing yellow fluorescent protein, Kanr Clontech GFP pEGFP-C1 Expressing enhanced green fluorescent protein, Kanr Clontech HGS/HGR-GFP pEGFP-C2 Expressing Chmp3; acts dominant-negative because of EGFP fusion, Kanr Tu et al. (85) VPS4A -GFP pEGFP-N1 Expressing a VPS4A-EGFP fusion protein, Kanr Tu et al.(85) VPS4A K173Q-GFP pEGFP-N1 Expressing a VPS4-EGFP fusion protein with point mutation in amino acid residue 173 of VPS4; acts as dominant-negative, Kanr Tu et al. (85) Bacterial vapA pET28a Expression vector for production of his6-tagged recombinant virulence-associated gene (VapA) from R. equi, lacking the secretion-mediating signal sequence, Kanr (8) aKanR, kanamycin resistant. .. order no. ALIX GUACCUCAGUCUAUACUUAtt s71368 CHMP3 GCUGGUUAAUGAAUGGUCAtt s83727 HRS/HGS AGUACAAGGUGGUCCAGGAtt s67517 TSG101 GCGUUAUCGAGGUAAUAUAtt s75514 VPS4A GAACAUACGGUGGAAUGAUtt s100252 VPS4B CGAAGAUUUGAGAAACGUAtt s73810 Mission siRNA universal negative control no. 1 = “scrambled siRNA” Sigma-Merck SIC001 Month XXXX Volume 0 Issue 0 10.1128/asmam.00010-2519 h ttp s: //j ou rn al s. as m .o rg /jo ur na l/a sm -a ni m al -m ic ro bi ol og y on 1 2 Fe br ua ry 2 02 6 by 2 40 0: 1a 00 :4 ba 5: 4c 42 :f 13 6: 57 9c :3 ae e: 68 f8
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