Mutagenesis:Article Title: Identification and Isolation of Genes Involved in Poly(?-Hydroxybutyrate) Biosynthesis in Azospirillum brasilense and Characterization of a phbC Mutant
Article Snippet: .. A. brasilense Sp7 (ATCC 29145) Wild-type strain 39 A. brasilense 7030 Sm r nitroso-guanidine-induced mutant of Sp7, lacking 115-MDa megaplasmid 12 A. lipoferum (ATCC 29708) Wild-type strain 39 A. amazonense (ATCC 35119) Wild-type strain 17 A. halopraeferens (ATCC 43709) Wild-type strain 30 A. irakense KBC1 Wild-type strain 15 A. doebereinerae GSF-71T Wild-type strain 10 A. brasilense phbC mutant Sp7 phbC ::Km mutant This study Escherichia coli DH5α hsdR17 endA1 thi-1 gyrA96 relA1 recA1 supE44 Δ lacU169 (φ80 lacZ ΔM15) 31 HB101 F − hsdS20 (r − B m − B ) recA13 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 supE44 31 S17.1 pro thi endA recA hsdR with RP4-2-Tc:: Mu -Km::Tn 7 integrated in chromosome, Sm r 32 Plasmids PUC18/19 Ap r , ColE1 replicon, lacZ , cloning vector 45 PLAFR3 Tc r , pLAFR1 derivative containing Hae II fragment of pUC8 33 pSUP202 Ap r , Tc r , Cm r , Col E1 replicon, mobilizable plasmid, suicide vector for A. brasilense 32 PRK2073 Sm r , Col E1 replicon, Tra + , Mob + , helper plasmid for triparental conjugations 11 PUCA800 pUC8 with Km r GenBlock from Puc-4K 20 P90 Tc r , pLAFR1 containing A. brasilense Sp7 p90 cosmid library 6 pP2EP5 Ap r , pUC19, containing 5-kb Eco RI- Pst I A. brasilense Sp7 phbC fragment This study pP13S2 Ap r , pUC18, containing 2-kb Sal I A. brasilense Sp7 phbA fragment This study pP13S8 Ap r , pUC18, containing 8-kb Sal I A. brasilense Sp7 phbB fragment This study pPEB1.2 Ap r , pUC18, containing 1.2-kb Eco RI- Bam HI A. brasilense Sp7 phbC fragment This study pPEBKm2.6 Ap r , Km r , pUC18, containing 2.6-kb Eco RI- Aat II A. brasilense Sp7 phbC fragment with Km r cassette inserted in Sma I site This study PSPEAKm Ap r , Km r pSUP202, containing 2.6-kb Eco RI- Aat II A. brasilense Sp7 phbC fragment with Km r cassette inserted in Sma I site This study Open in a separate window a Abbreviations: Tc, tetracycline; Ap, ampicillin; Km, kanamycin; Cm, cycloheximide, Sm, streptomycin. ..
Article Title: Identification and Isolation of Genes Involved in Poly(β-Hydroxybutyrate) Biosynthesis in Azospirillum brasilense and Characterization of a phbC Mutant
Article Snippet: .. A. brasilense Sp7 (ATCC 29145) Wild-type strain 39 A. brasilense 7030 Smr nitroso-guanidine-induced mutant of Sp7, lacking 115-MDa megaplasmid 12 A. lipoferum (ATCC 29708) Wild-type strain 39 A. amazonense (ATCC 35119) Wild-type strain 17 A. halopraeferens (ATCC 43709) Wild-type strain 30 A. irakense KBC1 Wild-type strain 15 A. doebereinerae GSF-71T Wild-type strain 10 A. brasilense phbC mutant Sp7 phbC::Km mutant This study Escherichia coli DH5 hsdR17 endA1 thi-1 gyrA96 relA1 recA1 supE44 lacU169( 80lacZ M15) 31 HB101 F hsdS20 (r B m B) recA13 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 supE44 31 S17.1 pro thi endA recA hsdR with RP4-2-Tc::Mu-Km::Tn7 integrated in chromosome, Smr 32 Plasmids PUC18/19 Apr, ColE1 replicon, lacZ, cloning vector 45 PLAFR3 Tcr, pLAFR1 derivative containing HaeII fragment of pUC8 33 pSUP202 Apr, Tcr, Cmr, ColE1 replicon, mobilizable plasmid, suicide vector for A. brasilense 32 PRK2073 Smr, ColE1 replicon, Tra , Mob , helper plasmid for triparental conjugations 11 PUCA800 pUC8 with Kmr GenBlock from Puc-4K 20 P90 Tcr, pLAFR1 containing A. brasilense Sp7 p90 cosmid library 6 pP2EP5 Apr, pUC19, containing 5-kb EcoRI-PstI A. brasilense Sp7 phbC fragment This study pP13S2 Apr, pUC18, containing 2-kb SalI A. brasilense Sp7 phbA fragment This study pP13S8 Apr, pUC18, containing 8-kb SalI A. brasilense Sp7 phbB fragment This study pPEB1.2 Apr, pUC18, containing 1.2-kb EcoRI-BamHI A. brasilense Sp7 phbC fragment This study pPEBKm2.6 Apr, Kmr, pUC18, containing 2.6-kb EcoRI-AatII A. brasilense Sp7 phbC fragment with Kmr cassette inserted in SmaI site This study PSPEAKm Apr, Kmr pSUP202, containing 2.6-kb EcoRI-AatII A. brasilense Sp7 phbC fragment with Kmr cassette inserted in SmaI site This study a Abbreviations: Tc, tetracycline; Ap, ampicillin; Km, kanamycin; Cm, cycloheximide, Sm, streptomycin. on A pril 7, 2015 by U N IV O F R H O D E IS LA N D http://aem .asm .org/ D ow nloaded from total DNA of A. brasilense Sp7 as the template in PCR, a 500-bp product, homologous to known phbC genes, was obtained. ..
Clone Assay:Article Title: Identification and Isolation of Genes Involved in Poly(?-Hydroxybutyrate) Biosynthesis in Azospirillum brasilense and Characterization of a phbC Mutant
Article Snippet: .. A. brasilense Sp7 (ATCC 29145) Wild-type strain 39 A. brasilense 7030 Sm r nitroso-guanidine-induced mutant of Sp7, lacking 115-MDa megaplasmid 12 A. lipoferum (ATCC 29708) Wild-type strain 39 A. amazonense (ATCC 35119) Wild-type strain 17 A. halopraeferens (ATCC 43709) Wild-type strain 30 A. irakense KBC1 Wild-type strain 15 A. doebereinerae GSF-71T Wild-type strain 10 A. brasilense phbC mutant Sp7 phbC ::Km mutant This study Escherichia coli DH5α hsdR17 endA1 thi-1 gyrA96 relA1 recA1 supE44 Δ lacU169 (φ80 lacZ ΔM15) 31 HB101 F − hsdS20 (r − B m − B ) recA13 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 supE44 31 S17.1 pro thi endA recA hsdR with RP4-2-Tc:: Mu -Km::Tn 7 integrated in chromosome, Sm r 32 Plasmids PUC18/19 Ap r , ColE1 replicon, lacZ , cloning vector 45 PLAFR3 Tc r , pLAFR1 derivative containing Hae II fragment of pUC8 33 pSUP202 Ap r , Tc r , Cm r , Col E1 replicon, mobilizable plasmid, suicide vector for A. brasilense 32 PRK2073 Sm r , Col E1 replicon, Tra + , Mob + , helper plasmid for triparental conjugations 11 PUCA800 pUC8 with Km r GenBlock from Puc-4K 20 P90 Tc r , pLAFR1 containing A. brasilense Sp7 p90 cosmid library 6 pP2EP5 Ap r , pUC19, containing 5-kb Eco RI- Pst I A. brasilense Sp7 phbC fragment This study pP13S2 Ap r , pUC18, containing 2-kb Sal I A. brasilense Sp7 phbA fragment This study pP13S8 Ap r , pUC18, containing 8-kb Sal I A. brasilense Sp7 phbB fragment This study pPEB1.2 Ap r , pUC18, containing 1.2-kb Eco RI- Bam HI A. brasilense Sp7 phbC fragment This study pPEBKm2.6 Ap r , Km r , pUC18, containing 2.6-kb Eco RI- Aat II A. brasilense Sp7 phbC fragment with Km r cassette inserted in Sma I site This study PSPEAKm Ap r , Km r pSUP202, containing 2.6-kb Eco RI- Aat II A. brasilense Sp7 phbC fragment with Km r cassette inserted in Sma I site This study Open in a separate window a Abbreviations: Tc, tetracycline; Ap, ampicillin; Km, kanamycin; Cm, cycloheximide, Sm, streptomycin. ..
Plasmid Preparation:Article Title: Identification and Isolation of Genes Involved in Poly(?-Hydroxybutyrate) Biosynthesis in Azospirillum brasilense and Characterization of a phbC Mutant
Article Snippet: .. A. brasilense Sp7 (ATCC 29145) Wild-type strain 39 A. brasilense 7030 Sm r nitroso-guanidine-induced mutant of Sp7, lacking 115-MDa megaplasmid 12 A. lipoferum (ATCC 29708) Wild-type strain 39 A. amazonense (ATCC 35119) Wild-type strain 17 A. halopraeferens (ATCC 43709) Wild-type strain 30 A. irakense KBC1 Wild-type strain 15 A. doebereinerae GSF-71T Wild-type strain 10 A. brasilense phbC mutant Sp7 phbC ::Km mutant This study Escherichia coli DH5α hsdR17 endA1 thi-1 gyrA96 relA1 recA1 supE44 Δ lacU169 (φ80 lacZ ΔM15) 31 HB101 F − hsdS20 (r − B m − B ) recA13 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 supE44 31 S17.1 pro thi endA recA hsdR with RP4-2-Tc:: Mu -Km::Tn 7 integrated in chromosome, Sm r 32 Plasmids PUC18/19 Ap r , ColE1 replicon, lacZ , cloning vector 45 PLAFR3 Tc r , pLAFR1 derivative containing Hae II fragment of pUC8 33 pSUP202 Ap r , Tc r , Cm r , Col E1 replicon, mobilizable plasmid, suicide vector for A. brasilense 32 PRK2073 Sm r , Col E1 replicon, Tra + , Mob + , helper plasmid for triparental conjugations 11 PUCA800 pUC8 with Km r GenBlock from Puc-4K 20 P90 Tc r , pLAFR1 containing A. brasilense Sp7 p90 cosmid library 6 pP2EP5 Ap r , pUC19, containing 5-kb Eco RI- Pst I A. brasilense Sp7 phbC fragment This study pP13S2 Ap r , pUC18, containing 2-kb Sal I A. brasilense Sp7 phbA fragment This study pP13S8 Ap r , pUC18, containing 8-kb Sal I A. brasilense Sp7 phbB fragment This study pPEB1.2 Ap r , pUC18, containing 1.2-kb Eco RI- Bam HI A. brasilense Sp7 phbC fragment This study pPEBKm2.6 Ap r , Km r , pUC18, containing 2.6-kb Eco RI- Aat II A. brasilense Sp7 phbC fragment with Km r cassette inserted in Sma I site This study PSPEAKm Ap r , Km r pSUP202, containing 2.6-kb Eco RI- Aat II A. brasilense Sp7 phbC fragment with Km r cassette inserted in Sma I site This study Open in a separate window a Abbreviations: Tc, tetracycline; Ap, ampicillin; Km, kanamycin; Cm, cycloheximide, Sm, streptomycin. ..
Article Title: Identification and Isolation of Genes Involved in Poly(β-Hydroxybutyrate) Biosynthesis in Azospirillum brasilense and Characterization of a phbC Mutant
Article Snippet: .. A. brasilense Sp7 (ATCC 29145) Wild-type strain 39 A. brasilense 7030 Smr nitroso-guanidine-induced mutant of Sp7, lacking 115-MDa megaplasmid 12 A. lipoferum (ATCC 29708) Wild-type strain 39 A. amazonense (ATCC 35119) Wild-type strain 17 A. halopraeferens (ATCC 43709) Wild-type strain 30 A. irakense KBC1 Wild-type strain 15 A. doebereinerae GSF-71T Wild-type strain 10 A. brasilense phbC mutant Sp7 phbC::Km mutant This study Escherichia coli DH5 hsdR17 endA1 thi-1 gyrA96 relA1 recA1 supE44 lacU169( 80lacZ M15) 31 HB101 F hsdS20 (r B m B) recA13 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 supE44 31 S17.1 pro thi endA recA hsdR with RP4-2-Tc::Mu-Km::Tn7 integrated in chromosome, Smr 32 Plasmids PUC18/19 Apr, ColE1 replicon, lacZ, cloning vector 45 PLAFR3 Tcr, pLAFR1 derivative containing HaeII fragment of pUC8 33 pSUP202 Apr, Tcr, Cmr, ColE1 replicon, mobilizable plasmid, suicide vector for A. brasilense 32 PRK2073 Smr, ColE1 replicon, Tra , Mob , helper plasmid for triparental conjugations 11 PUCA800 pUC8 with Kmr GenBlock from Puc-4K 20 P90 Tcr, pLAFR1 containing A. brasilense Sp7 p90 cosmid library 6 pP2EP5 Apr, pUC19, containing 5-kb EcoRI-PstI A. brasilense Sp7 phbC fragment This study pP13S2 Apr, pUC18, containing 2-kb SalI A. brasilense Sp7 phbA fragment This study pP13S8 Apr, pUC18, containing 8-kb SalI A. brasilense Sp7 phbB fragment This study pPEB1.2 Apr, pUC18, containing 1.2-kb EcoRI-BamHI A. brasilense Sp7 phbC fragment This study pPEBKm2.6 Apr, Kmr, pUC18, containing 2.6-kb EcoRI-AatII A. brasilense Sp7 phbC fragment with Kmr cassette inserted in SmaI site This study PSPEAKm Apr, Kmr pSUP202, containing 2.6-kb EcoRI-AatII A. brasilense Sp7 phbC fragment with Kmr cassette inserted in SmaI site This study a Abbreviations: Tc, tetracycline; Ap, ampicillin; Km, kanamycin; Cm, cycloheximide, Sm, streptomycin. on A pril 7, 2015 by U N IV O F R H O D E IS LA N D http://aem .asm .org/ D ow nloaded from total DNA of A. brasilense Sp7 as the template in PCR, a 500-bp product, homologous to known phbC genes, was obtained. ..
Cloning:Article Title: Identification and Isolation of Genes Involved in Poly(β-Hydroxybutyrate) Biosynthesis in Azospirillum brasilense and Characterization of a phbC Mutant
Article Snippet: .. A. brasilense Sp7 (ATCC 29145) Wild-type strain 39 A. brasilense 7030 Smr nitroso-guanidine-induced mutant of Sp7, lacking 115-MDa megaplasmid 12 A. lipoferum (ATCC 29708) Wild-type strain 39 A. amazonense (ATCC 35119) Wild-type strain 17 A. halopraeferens (ATCC 43709) Wild-type strain 30 A. irakense KBC1 Wild-type strain 15 A. doebereinerae GSF-71T Wild-type strain 10 A. brasilense phbC mutant Sp7 phbC::Km mutant This study Escherichia coli DH5 hsdR17 endA1 thi-1 gyrA96 relA1 recA1 supE44 lacU169( 80lacZ M15) 31 HB101 F hsdS20 (r B m B) recA13 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 supE44 31 S17.1 pro thi endA recA hsdR with RP4-2-Tc::Mu-Km::Tn7 integrated in chromosome, Smr 32 Plasmids PUC18/19 Apr, ColE1 replicon, lacZ, cloning vector 45 PLAFR3 Tcr, pLAFR1 derivative containing HaeII fragment of pUC8 33 pSUP202 Apr, Tcr, Cmr, ColE1 replicon, mobilizable plasmid, suicide vector for A. brasilense 32 PRK2073 Smr, ColE1 replicon, Tra , Mob , helper plasmid for triparental conjugations 11 PUCA800 pUC8 with Kmr GenBlock from Puc-4K 20 P90 Tcr, pLAFR1 containing A. brasilense Sp7 p90 cosmid library 6 pP2EP5 Apr, pUC19, containing 5-kb EcoRI-PstI A. brasilense Sp7 phbC fragment This study pP13S2 Apr, pUC18, containing 2-kb SalI A. brasilense Sp7 phbA fragment This study pP13S8 Apr, pUC18, containing 8-kb SalI A. brasilense Sp7 phbB fragment This study pPEB1.2 Apr, pUC18, containing 1.2-kb EcoRI-BamHI A. brasilense Sp7 phbC fragment This study pPEBKm2.6 Apr, Kmr, pUC18, containing 2.6-kb EcoRI-AatII A. brasilense Sp7 phbC fragment with Kmr cassette inserted in SmaI site This study PSPEAKm Apr, Kmr pSUP202, containing 2.6-kb EcoRI-AatII A. brasilense Sp7 phbC fragment with Kmr cassette inserted in SmaI site This study a Abbreviations: Tc, tetracycline; Ap, ampicillin; Km, kanamycin; Cm, cycloheximide, Sm, streptomycin. on A pril 7, 2015 by U N IV O F R H O D E IS LA N D http://aem .asm .org/ D ow nloaded from total DNA of A. brasilense Sp7 as the template in PCR, a 500-bp product, homologous to known phbC genes, was obtained. ..
Article Title: Identification and Isolation of Genes Involved in Poly(?-Hydroxybutyrate) Biosynthesis in Azospirillum brasilense and Characterization of a phbC Mutant
Article Snippet: .. A. brasilense Sp7 (ATCC 29145) Wild-type strain 39 A. brasilense 7030 Sm r nitroso-guanidine-induced mutant of Sp7, lacking 115-MDa megaplasmid 12 A. lipoferum (ATCC 29708) Wild-type strain 39 A. amazonense (ATCC 35119) Wild-type strain 17 A. halopraeferens (ATCC 43709) Wild-type strain 30 A. irakense KBC1 Wild-type strain 15 A. doebereinerae GSF-71T Wild-type strain 10 A. brasilense phbC mutant Sp7 phbC ::Km mutant This study Escherichia coli DH5α hsdR17 endA1 thi-1 gyrA96 relA1 recA1 supE44 Δ lacU169 (φ80 lacZ ΔM15) 31 HB101 F − hsdS20 (r − B m − B ) recA13 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 supE44 31 S17.1 pro thi endA recA hsdR with RP4-2-Tc:: Mu -Km::Tn 7 integrated in chromosome, Sm r 32 Plasmids PUC18/19 Ap r , ColE1 replicon, lacZ , cloning vector 45 PLAFR3 Tc r , pLAFR1 derivative containing Hae II fragment of pUC8 33 pSUP202 Ap r , Tc r , Cm r , Col E1 replicon, mobilizable plasmid, suicide vector for A. brasilense 32 PRK2073 Sm r , Col E1 replicon, Tra + , Mob + , helper plasmid for triparental conjugations 11 PUCA800 pUC8 with Km r GenBlock from Puc-4K 20 P90 Tc r , pLAFR1 containing A. brasilense Sp7 p90 cosmid library 6 pP2EP5 Ap r , pUC19, containing 5-kb Eco RI- Pst I A. brasilense Sp7 phbC fragment This study pP13S2 Ap r , pUC18, containing 2-kb Sal I A. brasilense Sp7 phbA fragment This study pP13S8 Ap r , pUC18, containing 8-kb Sal I A. brasilense Sp7 phbB fragment This study pPEB1.2 Ap r , pUC18, containing 1.2-kb Eco RI- Bam HI A. brasilense Sp7 phbC fragment This study pPEBKm2.6 Ap r , Km r , pUC18, containing 2.6-kb Eco RI- Aat II A. brasilense Sp7 phbC fragment with Km r cassette inserted in Sma I site This study PSPEAKm Ap r , Km r pSUP202, containing 2.6-kb Eco RI- Aat II A. brasilense Sp7 phbC fragment with Km r cassette inserted in Sma I site This study Open in a separate window a Abbreviations: Tc, tetracycline; Ap, ampicillin; Km, kanamycin; Cm, cycloheximide, Sm, streptomycin. ..
Polymerase Chain Reaction:Article Title: Identification and Isolation of Genes Involved in Poly(β-Hydroxybutyrate) Biosynthesis in Azospirillum brasilense and Characterization of a phbC Mutant
Article Snippet: .. A. brasilense Sp7 (ATCC 29145) Wild-type strain 39 A. brasilense 7030 Smr nitroso-guanidine-induced mutant of Sp7, lacking 115-MDa megaplasmid 12 A. lipoferum (ATCC 29708) Wild-type strain 39 A. amazonense (ATCC 35119) Wild-type strain 17 A. halopraeferens (ATCC 43709) Wild-type strain 30 A. irakense KBC1 Wild-type strain 15 A. doebereinerae GSF-71T Wild-type strain 10 A. brasilense phbC mutant Sp7 phbC::Km mutant This study Escherichia coli DH5 hsdR17 endA1 thi-1 gyrA96 relA1 recA1 supE44 lacU169( 80lacZ M15) 31 HB101 F hsdS20 (r B m B) recA13 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 supE44 31 S17.1 pro thi endA recA hsdR with RP4-2-Tc::Mu-Km::Tn7 integrated in chromosome, Smr 32 Plasmids PUC18/19 Apr, ColE1 replicon, lacZ, cloning vector 45 PLAFR3 Tcr, pLAFR1 derivative containing HaeII fragment of pUC8 33 pSUP202 Apr, Tcr, Cmr, ColE1 replicon, mobilizable plasmid, suicide vector for A. brasilense 32 PRK2073 Smr, ColE1 replicon, Tra , Mob , helper plasmid for triparental conjugations 11 PUCA800 pUC8 with Kmr GenBlock from Puc-4K 20 P90 Tcr, pLAFR1 containing A. brasilense Sp7 p90 cosmid library 6 pP2EP5 Apr, pUC19, containing 5-kb EcoRI-PstI A. brasilense Sp7 phbC fragment This study pP13S2 Apr, pUC18, containing 2-kb SalI A. brasilense Sp7 phbA fragment This study pP13S8 Apr, pUC18, containing 8-kb SalI A. brasilense Sp7 phbB fragment This study pPEB1.2 Apr, pUC18, containing 1.2-kb EcoRI-BamHI A. brasilense Sp7 phbC fragment This study pPEBKm2.6 Apr, Kmr, pUC18, containing 2.6-kb EcoRI-AatII A. brasilense Sp7 phbC fragment with Kmr cassette inserted in SmaI site This study PSPEAKm Apr, Kmr pSUP202, containing 2.6-kb EcoRI-AatII A. brasilense Sp7 phbC fragment with Kmr cassette inserted in SmaI site This study a Abbreviations: Tc, tetracycline; Ap, ampicillin; Km, kanamycin; Cm, cycloheximide, Sm, streptomycin. on A pril 7, 2015 by U N IV O F R H O D E IS LA N D http://aem .asm .org/ D ow nloaded from total DNA of A. brasilense Sp7 as the template in PCR, a 500-bp product, homologous to known phbC genes, was obtained. ..
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