Plasmid Preparation:Article Title: Transcription of the Gene Mediating Methicillin Resistance in Staphylococcus aureus ( mecA ) Is Corepressed but Not Coinduced by Cognate mecA and β-Lactamase Regulators
Article Snippet: .. Plasmids and strains Plasmid or strain Genotype or phenotype Remarks and reference Plasmids pUC19 Apr; 2.7 kb General cloning vector (35) pBluescript SK Apr; 3.0 kb General cloning vector (Stratagene) pRN5542 Cmr; 3 kb Staphylococcal cloning vector, derivative of pC194 (25) pJIM2246 Cmr; 6.0 kb Shuttle vector (27) pGO621 Apr; 5.2 kb mecR1-mecI ligated into puc19 at EcoRI-BamHI sites (this study) pGO621C Apr Cmr; 8.2 kb pGO621 with pSK265 ligated at BamHI site (this study) pGO630 Spr Tcr Emr ts; 16 kb Promoterless lacZ gene from E. coli fused to first 50 bp of mecA, intact mecR1-mecI ligated into pSK950 (31) pSK950 Emr Spr Tcr ts; 10.4 kb pCL84 with pE194ts at PstI site (24); integration vector pGO640 Apr; 5.2 kb PstI-EaeI fragment containing intact blaZ, blaR1, and truncated blaI from pCH2278 ligated into PstI-NotI sites of pBluescript KS pGO641 Apr Emr; 9.5 kb pE194 ligated into ClaI site of pGO640 (this study) pCH2278 Cmr; 6.2 kb 3.2-kb EcoRI-XhoI fragment containing bla locus ligated into EcoRISalI sites of pRN5542 (10) pCH1988 Cmr; 6.2 kb Same as pCH2278; contains linker insertion mutation in blaR1 (10) pGO630 R1 Spr Tcr Emr ts; 16 kb mecR1 mutant of pGO630 created by filling in ClaI site present in mecR1 (this study) pGO681 Cmr; 8.0 kb 2.0-kb BamHI-PstI fragment containing first 50 bp of mecA, intact mecR1, and mecI truncated at PstI site ligated into BamHI-PstI sites of pJIM2246 (this study) pGO682 Cmr; 8.8 kb 2.8-kb PstI-EaeI fragment containing intact blaZ, blaR1, and blaI truncated at EaeI site ligated into PstI-NotI sites of pJIM2246 (this study) pACT2 Apr; 8.1 kb Activation domain hybrid cloning vector (Clonetech) pAS1 Apr; 7.3 kb DNA-binding domain hybrid vector (Clonetech) S. aureus RN450 Cms Ems Gms Tcs bla ATCC 8235-4 RN4220 Cms Ems Gms Tcs bla Restriction-deficient mutagenized RN450 shuttle plasmid host (22) GO450TF Cms Ems Gms Tcs bla Provided by Tim Foster GO450TF::630 Cms Emr Gms Tcr bla geh GO450TF with integrated pGO630 (this study) GO450TF::630 I Cms Emr Gms Tcr bla geh mecR1 mecI mutant derivative of GO450TF::630 created by allelic replacement (this study) GO450TF::630 RI Cms Emr Gms Tcr bla geh mecR1 mecI derivative of GO450TF::630 created by allelic replacement (this study) .. DH5 80dlacZ M15 (lacZYA-argF)U169 deoR recA1 endA1 phoA hsdR17(r K m K) supE441 thi-1 gyrA96 relA91 glnV44 (7) Able C lac (LacZw ) (Kanr McrA McrB McrF Mrr hsdR(rK mK ) (F proAB laclqZ M15 Tn10(Tetr) (Stratagene)
Cloning:Article Title: Transcription of the Gene Mediating Methicillin Resistance in Staphylococcus aureus ( mecA ) Is Corepressed but Not Coinduced by Cognate mecA and β-Lactamase Regulators
Article Snippet: .. Plasmids and strains Plasmid or strain Genotype or phenotype Remarks and reference Plasmids pUC19 Apr; 2.7 kb General cloning vector (35) pBluescript SK Apr; 3.0 kb General cloning vector (Stratagene) pRN5542 Cmr; 3 kb Staphylococcal cloning vector, derivative of pC194 (25) pJIM2246 Cmr; 6.0 kb Shuttle vector (27) pGO621 Apr; 5.2 kb mecR1-mecI ligated into puc19 at EcoRI-BamHI sites (this study) pGO621C Apr Cmr; 8.2 kb pGO621 with pSK265 ligated at BamHI site (this study) pGO630 Spr Tcr Emr ts; 16 kb Promoterless lacZ gene from E. coli fused to first 50 bp of mecA, intact mecR1-mecI ligated into pSK950 (31) pSK950 Emr Spr Tcr ts; 10.4 kb pCL84 with pE194ts at PstI site (24); integration vector pGO640 Apr; 5.2 kb PstI-EaeI fragment containing intact blaZ, blaR1, and truncated blaI from pCH2278 ligated into PstI-NotI sites of pBluescript KS pGO641 Apr Emr; 9.5 kb pE194 ligated into ClaI site of pGO640 (this study) pCH2278 Cmr; 6.2 kb 3.2-kb EcoRI-XhoI fragment containing bla locus ligated into EcoRISalI sites of pRN5542 (10) pCH1988 Cmr; 6.2 kb Same as pCH2278; contains linker insertion mutation in blaR1 (10) pGO630 R1 Spr Tcr Emr ts; 16 kb mecR1 mutant of pGO630 created by filling in ClaI site present in mecR1 (this study) pGO681 Cmr; 8.0 kb 2.0-kb BamHI-PstI fragment containing first 50 bp of mecA, intact mecR1, and mecI truncated at PstI site ligated into BamHI-PstI sites of pJIM2246 (this study) pGO682 Cmr; 8.8 kb 2.8-kb PstI-EaeI fragment containing intact blaZ, blaR1, and blaI truncated at EaeI site ligated into PstI-NotI sites of pJIM2246 (this study) pACT2 Apr; 8.1 kb Activation domain hybrid cloning vector (Clonetech) pAS1 Apr; 7.3 kb DNA-binding domain hybrid vector (Clonetech) S. aureus RN450 Cms Ems Gms Tcs bla ATCC 8235-4 RN4220 Cms Ems Gms Tcs bla Restriction-deficient mutagenized RN450 shuttle plasmid host (22) GO450TF Cms Ems Gms Tcs bla Provided by Tim Foster GO450TF::630 Cms Emr Gms Tcr bla geh GO450TF with integrated pGO630 (this study) GO450TF::630 I Cms Emr Gms Tcr bla geh mecR1 mecI mutant derivative of GO450TF::630 created by allelic replacement (this study) GO450TF::630 RI Cms Emr Gms Tcr bla geh mecR1 mecI derivative of GO450TF::630 created by allelic replacement (this study) .. DH5 80dlacZ M15 (lacZYA-argF)U169 deoR recA1 endA1 phoA hsdR17(r K m K) supE441 thi-1 gyrA96 relA91 glnV44 (7) Able C lac (LacZw ) (Kanr McrA McrB McrF Mrr hsdR(rK mK ) (F proAB laclqZ M15 Tn10(Tetr) (Stratagene)
Mutagenesis:Article Title: Transcription of the Gene Mediating Methicillin Resistance in Staphylococcus aureus ( mecA ) Is Corepressed but Not Coinduced by Cognate mecA and β-Lactamase Regulators
Article Snippet: .. Plasmids and strains Plasmid or strain Genotype or phenotype Remarks and reference Plasmids pUC19 Apr; 2.7 kb General cloning vector (35) pBluescript SK Apr; 3.0 kb General cloning vector (Stratagene) pRN5542 Cmr; 3 kb Staphylococcal cloning vector, derivative of pC194 (25) pJIM2246 Cmr; 6.0 kb Shuttle vector (27) pGO621 Apr; 5.2 kb mecR1-mecI ligated into puc19 at EcoRI-BamHI sites (this study) pGO621C Apr Cmr; 8.2 kb pGO621 with pSK265 ligated at BamHI site (this study) pGO630 Spr Tcr Emr ts; 16 kb Promoterless lacZ gene from E. coli fused to first 50 bp of mecA, intact mecR1-mecI ligated into pSK950 (31) pSK950 Emr Spr Tcr ts; 10.4 kb pCL84 with pE194ts at PstI site (24); integration vector pGO640 Apr; 5.2 kb PstI-EaeI fragment containing intact blaZ, blaR1, and truncated blaI from pCH2278 ligated into PstI-NotI sites of pBluescript KS pGO641 Apr Emr; 9.5 kb pE194 ligated into ClaI site of pGO640 (this study) pCH2278 Cmr; 6.2 kb 3.2-kb EcoRI-XhoI fragment containing bla locus ligated into EcoRISalI sites of pRN5542 (10) pCH1988 Cmr; 6.2 kb Same as pCH2278; contains linker insertion mutation in blaR1 (10) pGO630 R1 Spr Tcr Emr ts; 16 kb mecR1 mutant of pGO630 created by filling in ClaI site present in mecR1 (this study) pGO681 Cmr; 8.0 kb 2.0-kb BamHI-PstI fragment containing first 50 bp of mecA, intact mecR1, and mecI truncated at PstI site ligated into BamHI-PstI sites of pJIM2246 (this study) pGO682 Cmr; 8.8 kb 2.8-kb PstI-EaeI fragment containing intact blaZ, blaR1, and blaI truncated at EaeI site ligated into PstI-NotI sites of pJIM2246 (this study) pACT2 Apr; 8.1 kb Activation domain hybrid cloning vector (Clonetech) pAS1 Apr; 7.3 kb DNA-binding domain hybrid vector (Clonetech) S. aureus RN450 Cms Ems Gms Tcs bla ATCC 8235-4 RN4220 Cms Ems Gms Tcs bla Restriction-deficient mutagenized RN450 shuttle plasmid host (22) GO450TF Cms Ems Gms Tcs bla Provided by Tim Foster GO450TF::630 Cms Emr Gms Tcr bla geh GO450TF with integrated pGO630 (this study) GO450TF::630 I Cms Emr Gms Tcr bla geh mecR1 mecI mutant derivative of GO450TF::630 created by allelic replacement (this study) GO450TF::630 RI Cms Emr Gms Tcr bla geh mecR1 mecI derivative of GO450TF::630 created by allelic replacement (this study) .. DH5 80dlacZ M15 (lacZYA-argF)U169 deoR recA1 endA1 phoA hsdR17(r K m K) supE441 thi-1 gyrA96 relA91 glnV44 (7) Able C lac (LacZw ) (Kanr McrA McrB McrF Mrr hsdR(rK mK ) (F proAB laclqZ M15 Tn10(Tetr) (Stratagene)
Activation Assay:Article Title: Transcription of the Gene Mediating Methicillin Resistance in Staphylococcus aureus ( mecA ) Is Corepressed but Not Coinduced by Cognate mecA and β-Lactamase Regulators
Article Snippet: .. Plasmids and strains Plasmid or strain Genotype or phenotype Remarks and reference Plasmids pUC19 Apr; 2.7 kb General cloning vector (35) pBluescript SK Apr; 3.0 kb General cloning vector (Stratagene) pRN5542 Cmr; 3 kb Staphylococcal cloning vector, derivative of pC194 (25) pJIM2246 Cmr; 6.0 kb Shuttle vector (27) pGO621 Apr; 5.2 kb mecR1-mecI ligated into puc19 at EcoRI-BamHI sites (this study) pGO621C Apr Cmr; 8.2 kb pGO621 with pSK265 ligated at BamHI site (this study) pGO630 Spr Tcr Emr ts; 16 kb Promoterless lacZ gene from E. coli fused to first 50 bp of mecA, intact mecR1-mecI ligated into pSK950 (31) pSK950 Emr Spr Tcr ts; 10.4 kb pCL84 with pE194ts at PstI site (24); integration vector pGO640 Apr; 5.2 kb PstI-EaeI fragment containing intact blaZ, blaR1, and truncated blaI from pCH2278 ligated into PstI-NotI sites of pBluescript KS pGO641 Apr Emr; 9.5 kb pE194 ligated into ClaI site of pGO640 (this study) pCH2278 Cmr; 6.2 kb 3.2-kb EcoRI-XhoI fragment containing bla locus ligated into EcoRISalI sites of pRN5542 (10) pCH1988 Cmr; 6.2 kb Same as pCH2278; contains linker insertion mutation in blaR1 (10) pGO630 R1 Spr Tcr Emr ts; 16 kb mecR1 mutant of pGO630 created by filling in ClaI site present in mecR1 (this study) pGO681 Cmr; 8.0 kb 2.0-kb BamHI-PstI fragment containing first 50 bp of mecA, intact mecR1, and mecI truncated at PstI site ligated into BamHI-PstI sites of pJIM2246 (this study) pGO682 Cmr; 8.8 kb 2.8-kb PstI-EaeI fragment containing intact blaZ, blaR1, and blaI truncated at EaeI site ligated into PstI-NotI sites of pJIM2246 (this study) pACT2 Apr; 8.1 kb Activation domain hybrid cloning vector (Clonetech) pAS1 Apr; 7.3 kb DNA-binding domain hybrid vector (Clonetech) S. aureus RN450 Cms Ems Gms Tcs bla ATCC 8235-4 RN4220 Cms Ems Gms Tcs bla Restriction-deficient mutagenized RN450 shuttle plasmid host (22) GO450TF Cms Ems Gms Tcs bla Provided by Tim Foster GO450TF::630 Cms Emr Gms Tcr bla geh GO450TF with integrated pGO630 (this study) GO450TF::630 I Cms Emr Gms Tcr bla geh mecR1 mecI mutant derivative of GO450TF::630 created by allelic replacement (this study) GO450TF::630 RI Cms Emr Gms Tcr bla geh mecR1 mecI derivative of GO450TF::630 created by allelic replacement (this study) .. DH5 80dlacZ M15 (lacZYA-argF)U169 deoR recA1 endA1 phoA hsdR17(r K m K) supE441 thi-1 gyrA96 relA91 glnV44 (7) Able C lac (LacZw ) (Kanr McrA McrB McrF Mrr hsdR(rK mK ) (F proAB laclqZ M15 Tn10(Tetr) (Stratagene)
|
ATCC is a verified supplier 
ATCC manufactures this product 