Cloning:Article Title: Functional Irreplaceability of Escherichia coli and Shewanella oneidensis OxyRs Is Critically Determined by Intrinsic Differences in Oligomerization
Article Snippet: .. E. coli strains MG1655 Wild type ATCC 700926 DH5α Host for cloning Lab stock BL21 Recombinant protein expression host strain Novagen WM3064 ΔdapA, donor strain for conjugation W. Metcalf, UIUC HG-EcOxyR ∆oxyR derived from MG1655 (1) S. oneidensis strains MR-1 Wild type ATCC 700550 HG1328 ∆oxyR derived from MR-1 (1) HG-OxyR-v ∆oxyR expressing one of all OxyR variants This study Plasmid pHGM01 Apr Gmr Cmr suicide vector (2) pHGEI01-PkatB For measuring PkatB activity (3) pET-28a Recombinant protein expression vector Novagen pET28a-OxyR-v All OxyR variants within pET-28a This study OxyR variants EcOxyR E. coli wild-type OxyR SoOxyR S. oneidensis wild-type OxyR Truncations OxyRDBD SoOxyR DBD domain only OxyRDBD EcOxyR DBD domain only SoOxyRΔXXX- SoOxyR lacking residues from XXX to the end Chimeras OxyRDBD-RD OxyR consisting of SoOxyR DBD and EcOxyR RD domains only OxyRDBD-RD OxyR consisting of EcOxyR DBD and SoOxyR RD domains only OxyRSXXXE OxyR consisting of SoOxyR sequence before residue XXX and EcOxyR sequence after residue XXX Point mutants EcOxyRC197S Residue 197 Cys replaced by Ser within EcOxyR SoOxyRC203S Residue 203 Cys replaced by Ser within SoOxyR SoOxyRL285A Residue 285 Leu replaced by Ala within SoOxyR SoOxyRT286A Residue 286 Thr replaced by Ala within SoOxyR SoOxyRF287A Residue 287 Phe replaced by Ala within SoOxyR SoOxyRR288A Residue 288 Arg replaced by Ala within SoOxyR SoOxyRT289A Residue 289 Thr replaced by Ala within SoOxyR SoOxyRL290A Residue 290 Leu replaced by Ala within SoOxyR SoOxyRG291A Residue 291 Gly replaced by Ala within SoOxyR SoOxyRR288E Residue 288 Arg replaced by Glu within SoOxyR SoOxyRR288E-L285A Point mutations for two residues SoOxyRR288E-T286A Point mutations for two residues SoOxyRR288E-F287A Point mutations for two residues SoOxyRR288E-T289A Point mutations for two residues SoOxyRR288E-L290A Point mutations for two residues SoOxyRR288E-G291A Point mutations for two residues References: 1. ..
Recombinant:Article Title: Functional Irreplaceability of Escherichia coli and Shewanella oneidensis OxyRs Is Critically Determined by Intrinsic Differences in Oligomerization
Article Snippet: .. E. coli strains MG1655 Wild type ATCC 700926 DH5α Host for cloning Lab stock BL21 Recombinant protein expression host strain Novagen WM3064 ΔdapA, donor strain for conjugation W. Metcalf, UIUC HG-EcOxyR ∆oxyR derived from MG1655 (1) S. oneidensis strains MR-1 Wild type ATCC 700550 HG1328 ∆oxyR derived from MR-1 (1) HG-OxyR-v ∆oxyR expressing one of all OxyR variants This study Plasmid pHGM01 Apr Gmr Cmr suicide vector (2) pHGEI01-PkatB For measuring PkatB activity (3) pET-28a Recombinant protein expression vector Novagen pET28a-OxyR-v All OxyR variants within pET-28a This study OxyR variants EcOxyR E. coli wild-type OxyR SoOxyR S. oneidensis wild-type OxyR Truncations OxyRDBD SoOxyR DBD domain only OxyRDBD EcOxyR DBD domain only SoOxyRΔXXX- SoOxyR lacking residues from XXX to the end Chimeras OxyRDBD-RD OxyR consisting of SoOxyR DBD and EcOxyR RD domains only OxyRDBD-RD OxyR consisting of EcOxyR DBD and SoOxyR RD domains only OxyRSXXXE OxyR consisting of SoOxyR sequence before residue XXX and EcOxyR sequence after residue XXX Point mutants EcOxyRC197S Residue 197 Cys replaced by Ser within EcOxyR SoOxyRC203S Residue 203 Cys replaced by Ser within SoOxyR SoOxyRL285A Residue 285 Leu replaced by Ala within SoOxyR SoOxyRT286A Residue 286 Thr replaced by Ala within SoOxyR SoOxyRF287A Residue 287 Phe replaced by Ala within SoOxyR SoOxyRR288A Residue 288 Arg replaced by Ala within SoOxyR SoOxyRT289A Residue 289 Thr replaced by Ala within SoOxyR SoOxyRL290A Residue 290 Leu replaced by Ala within SoOxyR SoOxyRG291A Residue 291 Gly replaced by Ala within SoOxyR SoOxyRR288E Residue 288 Arg replaced by Glu within SoOxyR SoOxyRR288E-L285A Point mutations for two residues SoOxyRR288E-T286A Point mutations for two residues SoOxyRR288E-F287A Point mutations for two residues SoOxyRR288E-T289A Point mutations for two residues SoOxyRR288E-L290A Point mutations for two residues SoOxyRR288E-G291A Point mutations for two residues References: 1. ..
Expressing:Article Title: Functional Irreplaceability of Escherichia coli and Shewanella oneidensis OxyRs Is Critically Determined by Intrinsic Differences in Oligomerization
Article Snippet: .. E. coli strains MG1655 Wild type ATCC 700926 DH5α Host for cloning Lab stock BL21 Recombinant protein expression host strain Novagen WM3064 ΔdapA, donor strain for conjugation W. Metcalf, UIUC HG-EcOxyR ∆oxyR derived from MG1655 (1) S. oneidensis strains MR-1 Wild type ATCC 700550 HG1328 ∆oxyR derived from MR-1 (1) HG-OxyR-v ∆oxyR expressing one of all OxyR variants This study Plasmid pHGM01 Apr Gmr Cmr suicide vector (2) pHGEI01-PkatB For measuring PkatB activity (3) pET-28a Recombinant protein expression vector Novagen pET28a-OxyR-v All OxyR variants within pET-28a This study OxyR variants EcOxyR E. coli wild-type OxyR SoOxyR S. oneidensis wild-type OxyR Truncations OxyRDBD SoOxyR DBD domain only OxyRDBD EcOxyR DBD domain only SoOxyRΔXXX- SoOxyR lacking residues from XXX to the end Chimeras OxyRDBD-RD OxyR consisting of SoOxyR DBD and EcOxyR RD domains only OxyRDBD-RD OxyR consisting of EcOxyR DBD and SoOxyR RD domains only OxyRSXXXE OxyR consisting of SoOxyR sequence before residue XXX and EcOxyR sequence after residue XXX Point mutants EcOxyRC197S Residue 197 Cys replaced by Ser within EcOxyR SoOxyRC203S Residue 203 Cys replaced by Ser within SoOxyR SoOxyRL285A Residue 285 Leu replaced by Ala within SoOxyR SoOxyRT286A Residue 286 Thr replaced by Ala within SoOxyR SoOxyRF287A Residue 287 Phe replaced by Ala within SoOxyR SoOxyRR288A Residue 288 Arg replaced by Ala within SoOxyR SoOxyRT289A Residue 289 Thr replaced by Ala within SoOxyR SoOxyRL290A Residue 290 Leu replaced by Ala within SoOxyR SoOxyRG291A Residue 291 Gly replaced by Ala within SoOxyR SoOxyRR288E Residue 288 Arg replaced by Glu within SoOxyR SoOxyRR288E-L285A Point mutations for two residues SoOxyRR288E-T286A Point mutations for two residues SoOxyRR288E-F287A Point mutations for two residues SoOxyRR288E-T289A Point mutations for two residues SoOxyRR288E-L290A Point mutations for two residues SoOxyRR288E-G291A Point mutations for two residues References: 1. ..
Conjugation Assay:Article Title: Functional Irreplaceability of Escherichia coli and Shewanella oneidensis OxyRs Is Critically Determined by Intrinsic Differences in Oligomerization
Article Snippet: .. E. coli strains MG1655 Wild type ATCC 700926 DH5α Host for cloning Lab stock BL21 Recombinant protein expression host strain Novagen WM3064 ΔdapA, donor strain for conjugation W. Metcalf, UIUC HG-EcOxyR ∆oxyR derived from MG1655 (1) S. oneidensis strains MR-1 Wild type ATCC 700550 HG1328 ∆oxyR derived from MR-1 (1) HG-OxyR-v ∆oxyR expressing one of all OxyR variants This study Plasmid pHGM01 Apr Gmr Cmr suicide vector (2) pHGEI01-PkatB For measuring PkatB activity (3) pET-28a Recombinant protein expression vector Novagen pET28a-OxyR-v All OxyR variants within pET-28a This study OxyR variants EcOxyR E. coli wild-type OxyR SoOxyR S. oneidensis wild-type OxyR Truncations OxyRDBD SoOxyR DBD domain only OxyRDBD EcOxyR DBD domain only SoOxyRΔXXX- SoOxyR lacking residues from XXX to the end Chimeras OxyRDBD-RD OxyR consisting of SoOxyR DBD and EcOxyR RD domains only OxyRDBD-RD OxyR consisting of EcOxyR DBD and SoOxyR RD domains only OxyRSXXXE OxyR consisting of SoOxyR sequence before residue XXX and EcOxyR sequence after residue XXX Point mutants EcOxyRC197S Residue 197 Cys replaced by Ser within EcOxyR SoOxyRC203S Residue 203 Cys replaced by Ser within SoOxyR SoOxyRL285A Residue 285 Leu replaced by Ala within SoOxyR SoOxyRT286A Residue 286 Thr replaced by Ala within SoOxyR SoOxyRF287A Residue 287 Phe replaced by Ala within SoOxyR SoOxyRR288A Residue 288 Arg replaced by Ala within SoOxyR SoOxyRT289A Residue 289 Thr replaced by Ala within SoOxyR SoOxyRL290A Residue 290 Leu replaced by Ala within SoOxyR SoOxyRG291A Residue 291 Gly replaced by Ala within SoOxyR SoOxyRR288E Residue 288 Arg replaced by Glu within SoOxyR SoOxyRR288E-L285A Point mutations for two residues SoOxyRR288E-T286A Point mutations for two residues SoOxyRR288E-F287A Point mutations for two residues SoOxyRR288E-T289A Point mutations for two residues SoOxyRR288E-L290A Point mutations for two residues SoOxyRR288E-G291A Point mutations for two residues References: 1. ..
Derivative Assay:Article Title: Functional Irreplaceability of Escherichia coli and Shewanella oneidensis OxyRs Is Critically Determined by Intrinsic Differences in Oligomerization
Article Snippet: .. E. coli strains MG1655 Wild type ATCC 700926 DH5α Host for cloning Lab stock BL21 Recombinant protein expression host strain Novagen WM3064 ΔdapA, donor strain for conjugation W. Metcalf, UIUC HG-EcOxyR ∆oxyR derived from MG1655 (1) S. oneidensis strains MR-1 Wild type ATCC 700550 HG1328 ∆oxyR derived from MR-1 (1) HG-OxyR-v ∆oxyR expressing one of all OxyR variants This study Plasmid pHGM01 Apr Gmr Cmr suicide vector (2) pHGEI01-PkatB For measuring PkatB activity (3) pET-28a Recombinant protein expression vector Novagen pET28a-OxyR-v All OxyR variants within pET-28a This study OxyR variants EcOxyR E. coli wild-type OxyR SoOxyR S. oneidensis wild-type OxyR Truncations OxyRDBD SoOxyR DBD domain only OxyRDBD EcOxyR DBD domain only SoOxyRΔXXX- SoOxyR lacking residues from XXX to the end Chimeras OxyRDBD-RD OxyR consisting of SoOxyR DBD and EcOxyR RD domains only OxyRDBD-RD OxyR consisting of EcOxyR DBD and SoOxyR RD domains only OxyRSXXXE OxyR consisting of SoOxyR sequence before residue XXX and EcOxyR sequence after residue XXX Point mutants EcOxyRC197S Residue 197 Cys replaced by Ser within EcOxyR SoOxyRC203S Residue 203 Cys replaced by Ser within SoOxyR SoOxyRL285A Residue 285 Leu replaced by Ala within SoOxyR SoOxyRT286A Residue 286 Thr replaced by Ala within SoOxyR SoOxyRF287A Residue 287 Phe replaced by Ala within SoOxyR SoOxyRR288A Residue 288 Arg replaced by Ala within SoOxyR SoOxyRT289A Residue 289 Thr replaced by Ala within SoOxyR SoOxyRL290A Residue 290 Leu replaced by Ala within SoOxyR SoOxyRG291A Residue 291 Gly replaced by Ala within SoOxyR SoOxyRR288E Residue 288 Arg replaced by Glu within SoOxyR SoOxyRR288E-L285A Point mutations for two residues SoOxyRR288E-T286A Point mutations for two residues SoOxyRR288E-F287A Point mutations for two residues SoOxyRR288E-T289A Point mutations for two residues SoOxyRR288E-L290A Point mutations for two residues SoOxyRR288E-G291A Point mutations for two residues References: 1. ..
Plasmid Preparation:Article Title: Functional Irreplaceability of Escherichia coli and Shewanella oneidensis OxyRs Is Critically Determined by Intrinsic Differences in Oligomerization
Article Snippet: .. E. coli strains MG1655 Wild type ATCC 700926 DH5α Host for cloning Lab stock BL21 Recombinant protein expression host strain Novagen WM3064 ΔdapA, donor strain for conjugation W. Metcalf, UIUC HG-EcOxyR ∆oxyR derived from MG1655 (1) S. oneidensis strains MR-1 Wild type ATCC 700550 HG1328 ∆oxyR derived from MR-1 (1) HG-OxyR-v ∆oxyR expressing one of all OxyR variants This study Plasmid pHGM01 Apr Gmr Cmr suicide vector (2) pHGEI01-PkatB For measuring PkatB activity (3) pET-28a Recombinant protein expression vector Novagen pET28a-OxyR-v All OxyR variants within pET-28a This study OxyR variants EcOxyR E. coli wild-type OxyR SoOxyR S. oneidensis wild-type OxyR Truncations OxyRDBD SoOxyR DBD domain only OxyRDBD EcOxyR DBD domain only SoOxyRΔXXX- SoOxyR lacking residues from XXX to the end Chimeras OxyRDBD-RD OxyR consisting of SoOxyR DBD and EcOxyR RD domains only OxyRDBD-RD OxyR consisting of EcOxyR DBD and SoOxyR RD domains only OxyRSXXXE OxyR consisting of SoOxyR sequence before residue XXX and EcOxyR sequence after residue XXX Point mutants EcOxyRC197S Residue 197 Cys replaced by Ser within EcOxyR SoOxyRC203S Residue 203 Cys replaced by Ser within SoOxyR SoOxyRL285A Residue 285 Leu replaced by Ala within SoOxyR SoOxyRT286A Residue 286 Thr replaced by Ala within SoOxyR SoOxyRF287A Residue 287 Phe replaced by Ala within SoOxyR SoOxyRR288A Residue 288 Arg replaced by Ala within SoOxyR SoOxyRT289A Residue 289 Thr replaced by Ala within SoOxyR SoOxyRL290A Residue 290 Leu replaced by Ala within SoOxyR SoOxyRG291A Residue 291 Gly replaced by Ala within SoOxyR SoOxyRR288E Residue 288 Arg replaced by Glu within SoOxyR SoOxyRR288E-L285A Point mutations for two residues SoOxyRR288E-T286A Point mutations for two residues SoOxyRR288E-F287A Point mutations for two residues SoOxyRR288E-T289A Point mutations for two residues SoOxyRR288E-L290A Point mutations for two residues SoOxyRR288E-G291A Point mutations for two residues References: 1. ..
Activity Assay:Article Title: Functional Irreplaceability of Escherichia coli and Shewanella oneidensis OxyRs Is Critically Determined by Intrinsic Differences in Oligomerization
Article Snippet: .. E. coli strains MG1655 Wild type ATCC 700926 DH5α Host for cloning Lab stock BL21 Recombinant protein expression host strain Novagen WM3064 ΔdapA, donor strain for conjugation W. Metcalf, UIUC HG-EcOxyR ∆oxyR derived from MG1655 (1) S. oneidensis strains MR-1 Wild type ATCC 700550 HG1328 ∆oxyR derived from MR-1 (1) HG-OxyR-v ∆oxyR expressing one of all OxyR variants This study Plasmid pHGM01 Apr Gmr Cmr suicide vector (2) pHGEI01-PkatB For measuring PkatB activity (3) pET-28a Recombinant protein expression vector Novagen pET28a-OxyR-v All OxyR variants within pET-28a This study OxyR variants EcOxyR E. coli wild-type OxyR SoOxyR S. oneidensis wild-type OxyR Truncations OxyRDBD SoOxyR DBD domain only OxyRDBD EcOxyR DBD domain only SoOxyRΔXXX- SoOxyR lacking residues from XXX to the end Chimeras OxyRDBD-RD OxyR consisting of SoOxyR DBD and EcOxyR RD domains only OxyRDBD-RD OxyR consisting of EcOxyR DBD and SoOxyR RD domains only OxyRSXXXE OxyR consisting of SoOxyR sequence before residue XXX and EcOxyR sequence after residue XXX Point mutants EcOxyRC197S Residue 197 Cys replaced by Ser within EcOxyR SoOxyRC203S Residue 203 Cys replaced by Ser within SoOxyR SoOxyRL285A Residue 285 Leu replaced by Ala within SoOxyR SoOxyRT286A Residue 286 Thr replaced by Ala within SoOxyR SoOxyRF287A Residue 287 Phe replaced by Ala within SoOxyR SoOxyRR288A Residue 288 Arg replaced by Ala within SoOxyR SoOxyRT289A Residue 289 Thr replaced by Ala within SoOxyR SoOxyRL290A Residue 290 Leu replaced by Ala within SoOxyR SoOxyRG291A Residue 291 Gly replaced by Ala within SoOxyR SoOxyRR288E Residue 288 Arg replaced by Glu within SoOxyR SoOxyRR288E-L285A Point mutations for two residues SoOxyRR288E-T286A Point mutations for two residues SoOxyRR288E-F287A Point mutations for two residues SoOxyRR288E-T289A Point mutations for two residues SoOxyRR288E-L290A Point mutations for two residues SoOxyRR288E-G291A Point mutations for two residues References: 1. ..
Sequencing:Article Title: Functional Irreplaceability of Escherichia coli and Shewanella oneidensis OxyRs Is Critically Determined by Intrinsic Differences in Oligomerization
Article Snippet: .. E. coli strains MG1655 Wild type ATCC 700926 DH5α Host for cloning Lab stock BL21 Recombinant protein expression host strain Novagen WM3064 ΔdapA, donor strain for conjugation W. Metcalf, UIUC HG-EcOxyR ∆oxyR derived from MG1655 (1) S. oneidensis strains MR-1 Wild type ATCC 700550 HG1328 ∆oxyR derived from MR-1 (1) HG-OxyR-v ∆oxyR expressing one of all OxyR variants This study Plasmid pHGM01 Apr Gmr Cmr suicide vector (2) pHGEI01-PkatB For measuring PkatB activity (3) pET-28a Recombinant protein expression vector Novagen pET28a-OxyR-v All OxyR variants within pET-28a This study OxyR variants EcOxyR E. coli wild-type OxyR SoOxyR S. oneidensis wild-type OxyR Truncations OxyRDBD SoOxyR DBD domain only OxyRDBD EcOxyR DBD domain only SoOxyRΔXXX- SoOxyR lacking residues from XXX to the end Chimeras OxyRDBD-RD OxyR consisting of SoOxyR DBD and EcOxyR RD domains only OxyRDBD-RD OxyR consisting of EcOxyR DBD and SoOxyR RD domains only OxyRSXXXE OxyR consisting of SoOxyR sequence before residue XXX and EcOxyR sequence after residue XXX Point mutants EcOxyRC197S Residue 197 Cys replaced by Ser within EcOxyR SoOxyRC203S Residue 203 Cys replaced by Ser within SoOxyR SoOxyRL285A Residue 285 Leu replaced by Ala within SoOxyR SoOxyRT286A Residue 286 Thr replaced by Ala within SoOxyR SoOxyRF287A Residue 287 Phe replaced by Ala within SoOxyR SoOxyRR288A Residue 288 Arg replaced by Ala within SoOxyR SoOxyRT289A Residue 289 Thr replaced by Ala within SoOxyR SoOxyRL290A Residue 290 Leu replaced by Ala within SoOxyR SoOxyRG291A Residue 291 Gly replaced by Ala within SoOxyR SoOxyRR288E Residue 288 Arg replaced by Glu within SoOxyR SoOxyRR288E-L285A Point mutations for two residues SoOxyRR288E-T286A Point mutations for two residues SoOxyRR288E-F287A Point mutations for two residues SoOxyRR288E-T289A Point mutations for two residues SoOxyRR288E-L290A Point mutations for two residues SoOxyRR288E-G291A Point mutations for two residues References: 1. ..
Residue:Article Title: Functional Irreplaceability of Escherichia coli and Shewanella oneidensis OxyRs Is Critically Determined by Intrinsic Differences in Oligomerization
Article Snippet: .. E. coli strains MG1655 Wild type ATCC 700926 DH5α Host for cloning Lab stock BL21 Recombinant protein expression host strain Novagen WM3064 ΔdapA, donor strain for conjugation W. Metcalf, UIUC HG-EcOxyR ∆oxyR derived from MG1655 (1) S. oneidensis strains MR-1 Wild type ATCC 700550 HG1328 ∆oxyR derived from MR-1 (1) HG-OxyR-v ∆oxyR expressing one of all OxyR variants This study Plasmid pHGM01 Apr Gmr Cmr suicide vector (2) pHGEI01-PkatB For measuring PkatB activity (3) pET-28a Recombinant protein expression vector Novagen pET28a-OxyR-v All OxyR variants within pET-28a This study OxyR variants EcOxyR E. coli wild-type OxyR SoOxyR S. oneidensis wild-type OxyR Truncations OxyRDBD SoOxyR DBD domain only OxyRDBD EcOxyR DBD domain only SoOxyRΔXXX- SoOxyR lacking residues from XXX to the end Chimeras OxyRDBD-RD OxyR consisting of SoOxyR DBD and EcOxyR RD domains only OxyRDBD-RD OxyR consisting of EcOxyR DBD and SoOxyR RD domains only OxyRSXXXE OxyR consisting of SoOxyR sequence before residue XXX and EcOxyR sequence after residue XXX Point mutants EcOxyRC197S Residue 197 Cys replaced by Ser within EcOxyR SoOxyRC203S Residue 203 Cys replaced by Ser within SoOxyR SoOxyRL285A Residue 285 Leu replaced by Ala within SoOxyR SoOxyRT286A Residue 286 Thr replaced by Ala within SoOxyR SoOxyRF287A Residue 287 Phe replaced by Ala within SoOxyR SoOxyRR288A Residue 288 Arg replaced by Ala within SoOxyR SoOxyRT289A Residue 289 Thr replaced by Ala within SoOxyR SoOxyRL290A Residue 290 Leu replaced by Ala within SoOxyR SoOxyRG291A Residue 291 Gly replaced by Ala within SoOxyR SoOxyRR288E Residue 288 Arg replaced by Glu within SoOxyR SoOxyRR288E-L285A Point mutations for two residues SoOxyRR288E-T286A Point mutations for two residues SoOxyRR288E-F287A Point mutations for two residues SoOxyRR288E-T289A Point mutations for two residues SoOxyRR288E-L290A Point mutations for two residues SoOxyRR288E-G291A Point mutations for two residues References: 1. ..
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