Construct:Article Title: The Cold-Induced Two-Component System CBO0366/CBO0365 Regulates Metabolic Pathways with Novel Roles in Group I Clostridium botulinum ATCC 3502 Cold Tolerance
Article Snippet: .. A single intron insertion was similarly demonstrated for the previously constructed cbo0365 mutant. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Strain or plasmid Relevant properties Source or reference Bacterial strains C. botulinum ATCC 3502 Wild-type parental strain ATCC a ATCC 3502 cbo0365 ::intron 48|49S Insertional disruption of cbo0365 at base 48 in sense orientation, erm 15 ATCC 3502 cbo0751 ::intron 254|255AS Insertional disruption of cbo0751 at base 254 in antisense orientation, erm This study ATCC 3502 cbo0753 ::intron 196|197AS Insertional disruption of cbo0753 at base 196 in antisense orientation, erm This study ATCC 3502 cbo1407 ::intron 428|429AS Insertional disruption of cbo1407 at base 428 in antisense orientation, erm This study ATCC 3502 cbo2525 ::intron 122|123AS Insertional disruption of cbo2525 at base 122 in antisense orientation, erm This study ATCC 3502 cbo2847 ::intron 509|510AS Insertional disruption of cbo2847 at base 509 in antisense orientation, erm This study ATCC 3502 cbo3199 ::intron 467|468AS Insertional disruption of cbo3199 at base 467 in antisense orientation, erm This study ATCC 3502 cbo3202 ::intron 167|168AS Insertional disruption of cbo3202 at base 167 in antisense orientation, erm This study E. coli CA434 Conjugation donor University of Nottingham Rosetta 2(DE3)/pLysS Recombinant expression host, pRARE2 encoding seven rare tRNAs, catP Novagen, Darmstadt, Germany Plasmids pMTL007C-E2 ClosTron plasmid, catP , L1.LtrB intron with ermB RAM, constitutive intron expression under fdx promoter University of Nottingham ( 19 ) pMTL007C-E2:: cbo0751 -254|255AS pMTL007C-E2 with L1.LtrB retargeted to base 254 of cbo0751 in antisense orientation This study pMTL007C-E2:: cbo0753 -196|197AS pMTL007C-E2 with L1.LtrB retargeted to base 196 of cbo0753 in antisense orientation This study pMTL007C-E2:: cbo1407 -428|429AS pMTL007C-E2 with L1.LtrB retargeted to base 428 of cbo1407 in antisense orientation This study pMTL007C-E2:: cbo2525 -122|123AS pMTL007C-E2 with L1.LtrB retargeted to base 122 of cbo2525 in antisense orientation This study pMTL007C-E2:: cbo2847 -509|510AS pMTL007C-E2 with L1.LtrB retargeted to base 509 of cbo2847 in antisense orientation This study pMTL007C-E2:: cbo3199 -467|468AS pMTL007C-E2 with L1.LtrB retargeted to base 467 of cbo3199 in antisense orientation This study pMTL007C-E2:: cbo3202 -167|168AS pMTL007C-E2 with L1.LtrB retargeted to base 167 of cbo3202 in antisense orientation This study pET-28b(+) Recombinant protein expression vector, kanR Novagen pET-28b(+)- cbo0365 -HIS pET-28b(+) harboring cbo0365 with N-terminal His 6 tag This study Open in a separate window a ATCC, American Type Culture Collection. ..
Mutagenesis:Article Title: The Cold-Induced Two-Component System CBO0366/CBO0365 Regulates Metabolic Pathways with Novel Roles in Group I Clostridium botulinum ATCC 3502 Cold Tolerance
Article Snippet: .. A single intron insertion was similarly demonstrated for the previously constructed cbo0365 mutant. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Strain or plasmid Relevant properties Source or reference Bacterial strains C. botulinum ATCC 3502 Wild-type parental strain ATCC a ATCC 3502 cbo0365 ::intron 48|49S Insertional disruption of cbo0365 at base 48 in sense orientation, erm 15 ATCC 3502 cbo0751 ::intron 254|255AS Insertional disruption of cbo0751 at base 254 in antisense orientation, erm This study ATCC 3502 cbo0753 ::intron 196|197AS Insertional disruption of cbo0753 at base 196 in antisense orientation, erm This study ATCC 3502 cbo1407 ::intron 428|429AS Insertional disruption of cbo1407 at base 428 in antisense orientation, erm This study ATCC 3502 cbo2525 ::intron 122|123AS Insertional disruption of cbo2525 at base 122 in antisense orientation, erm This study ATCC 3502 cbo2847 ::intron 509|510AS Insertional disruption of cbo2847 at base 509 in antisense orientation, erm This study ATCC 3502 cbo3199 ::intron 467|468AS Insertional disruption of cbo3199 at base 467 in antisense orientation, erm This study ATCC 3502 cbo3202 ::intron 167|168AS Insertional disruption of cbo3202 at base 167 in antisense orientation, erm This study E. coli CA434 Conjugation donor University of Nottingham Rosetta 2(DE3)/pLysS Recombinant expression host, pRARE2 encoding seven rare tRNAs, catP Novagen, Darmstadt, Germany Plasmids pMTL007C-E2 ClosTron plasmid, catP , L1.LtrB intron with ermB RAM, constitutive intron expression under fdx promoter University of Nottingham ( 19 ) pMTL007C-E2:: cbo0751 -254|255AS pMTL007C-E2 with L1.LtrB retargeted to base 254 of cbo0751 in antisense orientation This study pMTL007C-E2:: cbo0753 -196|197AS pMTL007C-E2 with L1.LtrB retargeted to base 196 of cbo0753 in antisense orientation This study pMTL007C-E2:: cbo1407 -428|429AS pMTL007C-E2 with L1.LtrB retargeted to base 428 of cbo1407 in antisense orientation This study pMTL007C-E2:: cbo2525 -122|123AS pMTL007C-E2 with L1.LtrB retargeted to base 122 of cbo2525 in antisense orientation This study pMTL007C-E2:: cbo2847 -509|510AS pMTL007C-E2 with L1.LtrB retargeted to base 509 of cbo2847 in antisense orientation This study pMTL007C-E2:: cbo3199 -467|468AS pMTL007C-E2 with L1.LtrB retargeted to base 467 of cbo3199 in antisense orientation This study pMTL007C-E2:: cbo3202 -167|168AS pMTL007C-E2 with L1.LtrB retargeted to base 167 of cbo3202 in antisense orientation This study pET-28b(+) Recombinant protein expression vector, kanR Novagen pET-28b(+)- cbo0365 -HIS pET-28b(+) harboring cbo0365 with N-terminal His 6 tag This study Open in a separate window a ATCC, American Type Culture Collection. ..
Plasmid Preparation:Article Title: The Cold-Induced Two-Component System CBO0366/CBO0365 Regulates Metabolic Pathways with Novel Roles in Group I Clostridium botulinum ATCC 3502 Cold Tolerance
Article Snippet: .. A single intron insertion was similarly demonstrated for the previously constructed cbo0365 mutant. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Strain or plasmid Relevant properties Source or reference Bacterial strains C. botulinum ATCC 3502 Wild-type parental strain ATCC a ATCC 3502 cbo0365 ::intron 48|49S Insertional disruption of cbo0365 at base 48 in sense orientation, erm 15 ATCC 3502 cbo0751 ::intron 254|255AS Insertional disruption of cbo0751 at base 254 in antisense orientation, erm This study ATCC 3502 cbo0753 ::intron 196|197AS Insertional disruption of cbo0753 at base 196 in antisense orientation, erm This study ATCC 3502 cbo1407 ::intron 428|429AS Insertional disruption of cbo1407 at base 428 in antisense orientation, erm This study ATCC 3502 cbo2525 ::intron 122|123AS Insertional disruption of cbo2525 at base 122 in antisense orientation, erm This study ATCC 3502 cbo2847 ::intron 509|510AS Insertional disruption of cbo2847 at base 509 in antisense orientation, erm This study ATCC 3502 cbo3199 ::intron 467|468AS Insertional disruption of cbo3199 at base 467 in antisense orientation, erm This study ATCC 3502 cbo3202 ::intron 167|168AS Insertional disruption of cbo3202 at base 167 in antisense orientation, erm This study E. coli CA434 Conjugation donor University of Nottingham Rosetta 2(DE3)/pLysS Recombinant expression host, pRARE2 encoding seven rare tRNAs, catP Novagen, Darmstadt, Germany Plasmids pMTL007C-E2 ClosTron plasmid, catP , L1.LtrB intron with ermB RAM, constitutive intron expression under fdx promoter University of Nottingham ( 19 ) pMTL007C-E2:: cbo0751 -254|255AS pMTL007C-E2 with L1.LtrB retargeted to base 254 of cbo0751 in antisense orientation This study pMTL007C-E2:: cbo0753 -196|197AS pMTL007C-E2 with L1.LtrB retargeted to base 196 of cbo0753 in antisense orientation This study pMTL007C-E2:: cbo1407 -428|429AS pMTL007C-E2 with L1.LtrB retargeted to base 428 of cbo1407 in antisense orientation This study pMTL007C-E2:: cbo2525 -122|123AS pMTL007C-E2 with L1.LtrB retargeted to base 122 of cbo2525 in antisense orientation This study pMTL007C-E2:: cbo2847 -509|510AS pMTL007C-E2 with L1.LtrB retargeted to base 509 of cbo2847 in antisense orientation This study pMTL007C-E2:: cbo3199 -467|468AS pMTL007C-E2 with L1.LtrB retargeted to base 467 of cbo3199 in antisense orientation This study pMTL007C-E2:: cbo3202 -167|168AS pMTL007C-E2 with L1.LtrB retargeted to base 167 of cbo3202 in antisense orientation This study pET-28b(+) Recombinant protein expression vector, kanR Novagen pET-28b(+)- cbo0365 -HIS pET-28b(+) harboring cbo0365 with N-terminal His 6 tag This study Open in a separate window a ATCC, American Type Culture Collection. ..
Disruption:Article Title: The Cold-Induced Two-Component System CBO0366/CBO0365 Regulates Metabolic Pathways with Novel Roles in Group I Clostridium botulinum ATCC 3502 Cold Tolerance
Article Snippet: .. A single intron insertion was similarly demonstrated for the previously constructed cbo0365 mutant. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Strain or plasmid Relevant properties Source or reference Bacterial strains C. botulinum ATCC 3502 Wild-type parental strain ATCC a ATCC 3502 cbo0365 ::intron 48|49S Insertional disruption of cbo0365 at base 48 in sense orientation, erm 15 ATCC 3502 cbo0751 ::intron 254|255AS Insertional disruption of cbo0751 at base 254 in antisense orientation, erm This study ATCC 3502 cbo0753 ::intron 196|197AS Insertional disruption of cbo0753 at base 196 in antisense orientation, erm This study ATCC 3502 cbo1407 ::intron 428|429AS Insertional disruption of cbo1407 at base 428 in antisense orientation, erm This study ATCC 3502 cbo2525 ::intron 122|123AS Insertional disruption of cbo2525 at base 122 in antisense orientation, erm This study ATCC 3502 cbo2847 ::intron 509|510AS Insertional disruption of cbo2847 at base 509 in antisense orientation, erm This study ATCC 3502 cbo3199 ::intron 467|468AS Insertional disruption of cbo3199 at base 467 in antisense orientation, erm This study ATCC 3502 cbo3202 ::intron 167|168AS Insertional disruption of cbo3202 at base 167 in antisense orientation, erm This study E. coli CA434 Conjugation donor University of Nottingham Rosetta 2(DE3)/pLysS Recombinant expression host, pRARE2 encoding seven rare tRNAs, catP Novagen, Darmstadt, Germany Plasmids pMTL007C-E2 ClosTron plasmid, catP , L1.LtrB intron with ermB RAM, constitutive intron expression under fdx promoter University of Nottingham ( 19 ) pMTL007C-E2:: cbo0751 -254|255AS pMTL007C-E2 with L1.LtrB retargeted to base 254 of cbo0751 in antisense orientation This study pMTL007C-E2:: cbo0753 -196|197AS pMTL007C-E2 with L1.LtrB retargeted to base 196 of cbo0753 in antisense orientation This study pMTL007C-E2:: cbo1407 -428|429AS pMTL007C-E2 with L1.LtrB retargeted to base 428 of cbo1407 in antisense orientation This study pMTL007C-E2:: cbo2525 -122|123AS pMTL007C-E2 with L1.LtrB retargeted to base 122 of cbo2525 in antisense orientation This study pMTL007C-E2:: cbo2847 -509|510AS pMTL007C-E2 with L1.LtrB retargeted to base 509 of cbo2847 in antisense orientation This study pMTL007C-E2:: cbo3199 -467|468AS pMTL007C-E2 with L1.LtrB retargeted to base 467 of cbo3199 in antisense orientation This study pMTL007C-E2:: cbo3202 -167|168AS pMTL007C-E2 with L1.LtrB retargeted to base 167 of cbo3202 in antisense orientation This study pET-28b(+) Recombinant protein expression vector, kanR Novagen pET-28b(+)- cbo0365 -HIS pET-28b(+) harboring cbo0365 with N-terminal His 6 tag This study Open in a separate window a ATCC, American Type Culture Collection. ..
Conjugation Assay:Article Title: The Cold-Induced Two-Component System CBO0366/CBO0365 Regulates Metabolic Pathways with Novel Roles in Group I Clostridium botulinum ATCC 3502 Cold Tolerance
Article Snippet: .. A single intron insertion was similarly demonstrated for the previously constructed cbo0365 mutant. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Strain or plasmid Relevant properties Source or reference Bacterial strains C. botulinum ATCC 3502 Wild-type parental strain ATCC a ATCC 3502 cbo0365 ::intron 48|49S Insertional disruption of cbo0365 at base 48 in sense orientation, erm 15 ATCC 3502 cbo0751 ::intron 254|255AS Insertional disruption of cbo0751 at base 254 in antisense orientation, erm This study ATCC 3502 cbo0753 ::intron 196|197AS Insertional disruption of cbo0753 at base 196 in antisense orientation, erm This study ATCC 3502 cbo1407 ::intron 428|429AS Insertional disruption of cbo1407 at base 428 in antisense orientation, erm This study ATCC 3502 cbo2525 ::intron 122|123AS Insertional disruption of cbo2525 at base 122 in antisense orientation, erm This study ATCC 3502 cbo2847 ::intron 509|510AS Insertional disruption of cbo2847 at base 509 in antisense orientation, erm This study ATCC 3502 cbo3199 ::intron 467|468AS Insertional disruption of cbo3199 at base 467 in antisense orientation, erm This study ATCC 3502 cbo3202 ::intron 167|168AS Insertional disruption of cbo3202 at base 167 in antisense orientation, erm This study E. coli CA434 Conjugation donor University of Nottingham Rosetta 2(DE3)/pLysS Recombinant expression host, pRARE2 encoding seven rare tRNAs, catP Novagen, Darmstadt, Germany Plasmids pMTL007C-E2 ClosTron plasmid, catP , L1.LtrB intron with ermB RAM, constitutive intron expression under fdx promoter University of Nottingham ( 19 ) pMTL007C-E2:: cbo0751 -254|255AS pMTL007C-E2 with L1.LtrB retargeted to base 254 of cbo0751 in antisense orientation This study pMTL007C-E2:: cbo0753 -196|197AS pMTL007C-E2 with L1.LtrB retargeted to base 196 of cbo0753 in antisense orientation This study pMTL007C-E2:: cbo1407 -428|429AS pMTL007C-E2 with L1.LtrB retargeted to base 428 of cbo1407 in antisense orientation This study pMTL007C-E2:: cbo2525 -122|123AS pMTL007C-E2 with L1.LtrB retargeted to base 122 of cbo2525 in antisense orientation This study pMTL007C-E2:: cbo2847 -509|510AS pMTL007C-E2 with L1.LtrB retargeted to base 509 of cbo2847 in antisense orientation This study pMTL007C-E2:: cbo3199 -467|468AS pMTL007C-E2 with L1.LtrB retargeted to base 467 of cbo3199 in antisense orientation This study pMTL007C-E2:: cbo3202 -167|168AS pMTL007C-E2 with L1.LtrB retargeted to base 167 of cbo3202 in antisense orientation This study pET-28b(+) Recombinant protein expression vector, kanR Novagen pET-28b(+)- cbo0365 -HIS pET-28b(+) harboring cbo0365 with N-terminal His 6 tag This study Open in a separate window a ATCC, American Type Culture Collection. ..
Recombinant:Article Title: The Cold-Induced Two-Component System CBO0366/CBO0365 Regulates Metabolic Pathways with Novel Roles in Group I Clostridium botulinum ATCC 3502 Cold Tolerance
Article Snippet: .. A single intron insertion was similarly demonstrated for the previously constructed cbo0365 mutant. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Strain or plasmid Relevant properties Source or reference Bacterial strains C. botulinum ATCC 3502 Wild-type parental strain ATCC a ATCC 3502 cbo0365 ::intron 48|49S Insertional disruption of cbo0365 at base 48 in sense orientation, erm 15 ATCC 3502 cbo0751 ::intron 254|255AS Insertional disruption of cbo0751 at base 254 in antisense orientation, erm This study ATCC 3502 cbo0753 ::intron 196|197AS Insertional disruption of cbo0753 at base 196 in antisense orientation, erm This study ATCC 3502 cbo1407 ::intron 428|429AS Insertional disruption of cbo1407 at base 428 in antisense orientation, erm This study ATCC 3502 cbo2525 ::intron 122|123AS Insertional disruption of cbo2525 at base 122 in antisense orientation, erm This study ATCC 3502 cbo2847 ::intron 509|510AS Insertional disruption of cbo2847 at base 509 in antisense orientation, erm This study ATCC 3502 cbo3199 ::intron 467|468AS Insertional disruption of cbo3199 at base 467 in antisense orientation, erm This study ATCC 3502 cbo3202 ::intron 167|168AS Insertional disruption of cbo3202 at base 167 in antisense orientation, erm This study E. coli CA434 Conjugation donor University of Nottingham Rosetta 2(DE3)/pLysS Recombinant expression host, pRARE2 encoding seven rare tRNAs, catP Novagen, Darmstadt, Germany Plasmids pMTL007C-E2 ClosTron plasmid, catP , L1.LtrB intron with ermB RAM, constitutive intron expression under fdx promoter University of Nottingham ( 19 ) pMTL007C-E2:: cbo0751 -254|255AS pMTL007C-E2 with L1.LtrB retargeted to base 254 of cbo0751 in antisense orientation This study pMTL007C-E2:: cbo0753 -196|197AS pMTL007C-E2 with L1.LtrB retargeted to base 196 of cbo0753 in antisense orientation This study pMTL007C-E2:: cbo1407 -428|429AS pMTL007C-E2 with L1.LtrB retargeted to base 428 of cbo1407 in antisense orientation This study pMTL007C-E2:: cbo2525 -122|123AS pMTL007C-E2 with L1.LtrB retargeted to base 122 of cbo2525 in antisense orientation This study pMTL007C-E2:: cbo2847 -509|510AS pMTL007C-E2 with L1.LtrB retargeted to base 509 of cbo2847 in antisense orientation This study pMTL007C-E2:: cbo3199 -467|468AS pMTL007C-E2 with L1.LtrB retargeted to base 467 of cbo3199 in antisense orientation This study pMTL007C-E2:: cbo3202 -167|168AS pMTL007C-E2 with L1.LtrB retargeted to base 167 of cbo3202 in antisense orientation This study pET-28b(+) Recombinant protein expression vector, kanR Novagen pET-28b(+)- cbo0365 -HIS pET-28b(+) harboring cbo0365 with N-terminal His 6 tag This study Open in a separate window a ATCC, American Type Culture Collection. ..
Expressing:Article Title: The Cold-Induced Two-Component System CBO0366/CBO0365 Regulates Metabolic Pathways with Novel Roles in Group I Clostridium botulinum ATCC 3502 Cold Tolerance
Article Snippet: .. A single intron insertion was similarly demonstrated for the previously constructed cbo0365 mutant. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Strain or plasmid Relevant properties Source or reference Bacterial strains C. botulinum ATCC 3502 Wild-type parental strain ATCC a ATCC 3502 cbo0365 ::intron 48|49S Insertional disruption of cbo0365 at base 48 in sense orientation, erm 15 ATCC 3502 cbo0751 ::intron 254|255AS Insertional disruption of cbo0751 at base 254 in antisense orientation, erm This study ATCC 3502 cbo0753 ::intron 196|197AS Insertional disruption of cbo0753 at base 196 in antisense orientation, erm This study ATCC 3502 cbo1407 ::intron 428|429AS Insertional disruption of cbo1407 at base 428 in antisense orientation, erm This study ATCC 3502 cbo2525 ::intron 122|123AS Insertional disruption of cbo2525 at base 122 in antisense orientation, erm This study ATCC 3502 cbo2847 ::intron 509|510AS Insertional disruption of cbo2847 at base 509 in antisense orientation, erm This study ATCC 3502 cbo3199 ::intron 467|468AS Insertional disruption of cbo3199 at base 467 in antisense orientation, erm This study ATCC 3502 cbo3202 ::intron 167|168AS Insertional disruption of cbo3202 at base 167 in antisense orientation, erm This study E. coli CA434 Conjugation donor University of Nottingham Rosetta 2(DE3)/pLysS Recombinant expression host, pRARE2 encoding seven rare tRNAs, catP Novagen, Darmstadt, Germany Plasmids pMTL007C-E2 ClosTron plasmid, catP , L1.LtrB intron with ermB RAM, constitutive intron expression under fdx promoter University of Nottingham ( 19 ) pMTL007C-E2:: cbo0751 -254|255AS pMTL007C-E2 with L1.LtrB retargeted to base 254 of cbo0751 in antisense orientation This study pMTL007C-E2:: cbo0753 -196|197AS pMTL007C-E2 with L1.LtrB retargeted to base 196 of cbo0753 in antisense orientation This study pMTL007C-E2:: cbo1407 -428|429AS pMTL007C-E2 with L1.LtrB retargeted to base 428 of cbo1407 in antisense orientation This study pMTL007C-E2:: cbo2525 -122|123AS pMTL007C-E2 with L1.LtrB retargeted to base 122 of cbo2525 in antisense orientation This study pMTL007C-E2:: cbo2847 -509|510AS pMTL007C-E2 with L1.LtrB retargeted to base 509 of cbo2847 in antisense orientation This study pMTL007C-E2:: cbo3199 -467|468AS pMTL007C-E2 with L1.LtrB retargeted to base 467 of cbo3199 in antisense orientation This study pMTL007C-E2:: cbo3202 -167|168AS pMTL007C-E2 with L1.LtrB retargeted to base 167 of cbo3202 in antisense orientation This study pET-28b(+) Recombinant protein expression vector, kanR Novagen pET-28b(+)- cbo0365 -HIS pET-28b(+) harboring cbo0365 with N-terminal His 6 tag This study Open in a separate window a ATCC, American Type Culture Collection. ..
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