Journal:

Article Title: Metabolic Roles of a Rhodobacter sphaeroides Member of the ? 32 Family

doi:

Figure Lengend Snippet: Bacterial strains and plasmids

Article Snippet: Plating efficiency of phage RS1 was determined with aerobically grown cells ( 6 ). table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Strain or plasmid Relevant feature(s) Source or reference R. sphaeroides 2.4.1 Wild type Laboratory strain RpoH1 Δ rpoH1 ::ΩSp r derivative of 2.4.1 This work RpoH26 Δ rpoH1 ::ΩSp r derivative of 2.4.1 with pPJR19 integrated This work E. coli DH5α Strain used for plasmid maintenance 3 S17-1 Conjugative donor for mating 36 CAG12517 MC4100 (R40) Δ rpoH30 :: kan , suppressor tRNA 44 Plasmids pGEM-7Zf(+) Ap r Promega pSUP202 Ap r Tc r Cm r R. sphaeroides suicide plasmid 36 pHP45Ω Ap r Sp r source of 2-kb ΩSp r cartridge 31 pPJR18 4.3-kb Eco RI restriction fragment of rpoH in pGEM-7Zf(+) This work pPRJ19 pPJR18 lacking 3-kb Sma I restriction fragment This work pPJR26 pPJR19 Sty I restriction fragment replaced by ΩSp r cartridge This work pPJR29 rpoH region from pPRJ26 in pSUP202 This work Open in a separate window Bacterial strains and plasmids Tellurite sensitivity was tested by diluting aerobically grown cells (∼10 9 cells/ml) sufficiently to produce ∼100 colonies per plate ( 23 ).

Techniques: Plasmid Preparation

Journal: Bioscience, biotechnology, and biochemistry

Article Title: Main polyol dehydrogenase of Gluconobacter suboxydans IFO 3255, membrane-bound D-sorbitol dehydrogenase, that needs product of upstream gene, sldB, for activity.

doi: 10.1271/bbb.66.2314

Figure Lengend Snippet: Fig. 1. Genetic Structure of the Plasmids for Disrupting the sldA and sldB Genes. A, Genetic structures of pSUP202-sldA::Km and pSUP202-sldB::Gm-ptac. B, The detailed structure of the 0.9-kb SalI fragment in plasmid pTNB148. Tc, tetracycline resistance; Km, kanamycin resistance; Gm, gentamicin; Ap, ampicillin; Cm, chloramphenicol resistance; mob, mobilization; arrows, the transcriptional orientation of each gene.

Article Snippet: [traD36, proAB +, lacI q, lacZDM15] E. coli HB101 F„, hsdS20, recA13, ara-14, proA2, lacY1, galK2, rpsL20 (str), xyl-5, mtl-1, sup E44, leuB6, thi-1 Plasmids pVK100 RK2 (IncP) derivative 23-kb Kmr Tcr cos ATCCc pRK2013 RK2 (IncP) derivative 45-kb Kmr Tra mob ATCC pSUP202 Apr Tcr Cmr mob 12) pCRII Apr Kmr PCR cloning vector pUC18 Apr E. coli cloning vector pUC-4K Kmr gene cassette pUC-8G Gmr gene cassette This study pUC-pA pUC18 carrying pA This study pCHR65 pBR322-based plasmid with Gmr gene 16) pDR540 Apr tac promoter vector pVKSLP pVK100 carrying 6.2-kb PstI fragment carrying sldA and sldB genes This study pTNB136 pVK100 carrying modiˆed sldA gene This study pTNB148 pVK100 carrying modiˆed sldB gene This study a Drug resistance: Tc, tetracycline; Km, kanamycin; Gm, gentamicin; Ap, ampicillin; Cm, chloramphenicol. b IFO: Institute for Fermentation, Osaka c ATCC: American Type Culture Collection Gene Disruption of D-Sorbitol Dehydrogenase of Gluconobacter unit with dehydrogenating activity, analogous to the membrane-bound D-glucose dehydrogenase consisting of two domains described by Yamada et al.11) These researchers proposed a topological model of E. coli D-glucose dehydrogenase, with an N-terminal hydrophobic domain of about 150 amino acid residues and a dehydrogenase domain of about 650 amino acid residues.

Techniques: Plasmid Preparation

Journal:

Article Title: Importance of Rhodospirillum rubrum H + -Pyrophosphatase under Low-Energy Conditions

doi: 10.1128/JB.186.19.6651-6655.2004

Figure Lengend Snippet: Bacterial strains and plasmids

Article Snippet: Wild type Rhodomicrobium vannielii DSM 162 Wild type Escherichia coli S17-1 pro thi hsdR hsdM + recA [RP4-2 Tc::Mu-Kan::Tn 7 (Tp r Sm r ) Tra + ] 22 Plasmids pCRII 3.9-kb cloning vector, Ap r Kan r Invitrogen pCR-Hpp pCRII 1.9-kb fragment of hpp This work pCR-Hpp-K pCRII- hpp-960 ::Kan This work pUC4K pUC18 derivative, Ap r Kan r 30 pSUP202 Suicide vector, Mob + Cm r Ap r Tc r 22 pSUP-Hpp-K pSUP202 carrying 1.9-kb fragment of hpp-960 ::Kan, Ap r Cm r This work pBBR1MCS-5 4.8-kb cloning vector, Mob + Gm r 15 pBBR-Hpp-Rr pBBR1MCS-5 ( hpp ) This work Open in a separate window a hpp , R. rubrum wild-type H + PPase gene.

Techniques: Plasmid Preparation, Clone Assay

Journal:

Article Title: Identification and Isolation of Genes Involved in Poly(?-Hydroxybutyrate) Biosynthesis in Azospirillum brasilense and Characterization of a phbC Mutant

doi: 10.1128/AEM.68.6.2943-2949.2002

Figure Lengend Snippet: Bacterial strains and plasmids used in this study

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.

Techniques: Plasmid Preparation, Mutagenesis, Clone Assay

Journal:

Article Title: Identification and Isolation of Genes Involved in Poly(?-Hydroxybutyrate) Biosynthesis in Azospirillum brasilense and Characterization of a phbC Mutant

doi: 10.1128/AEM.68.6.2943-2949.2002

Figure Lengend Snippet: Multiple alignment of the deduced amino acid sequence of phbC of A. brasilense (accession no. AF353205) with corresponding sequences of phbC from R. rubrum (accession no. CAB65395), A. caulinodans (accession no. CAA06928), S. meliloti (accession no. AAC61899), M. loti (accession no. BAB48418), and R. eutrophus (accession no. AAB24910). Three levels of similarity are shown according to the default settings of GeneDoc.

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.

Techniques: Sequencing

Journal:

Article Title: Identification and Isolation of Genes Involved in Poly(?-Hydroxybutyrate) Biosynthesis in Azospirillum brasilense and Characterization of a phbC Mutant

doi: 10.1128/AEM.68.6.2943-2949.2002

Figure Lengend Snippet: Physical map of A. brasilense Sp7 phbABC and fadE region. Open arrows indicate the locations and directions of transcription of phbC, phbA, phbB, and fadE. The triangle represents the location of the insertion of the kanamycin resistance cassette in the phbC mutant strain. E, EcoRI; B, BamHI; S, SmaI.

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.

Techniques: Mutagenesis

Journal:

Article Title: Identification and Isolation of Genes Involved in Poly(?-Hydroxybutyrate) Biosynthesis in Azospirillum brasilense and Characterization of a phbC Mutant

doi: 10.1128/AEM.68.6.2943-2949.2002

Figure Lengend Snippet: Southern blot hybridization using a DIG-labeled 500-bp long PCR-generated probe representing an internal region of the A. brasilense Sp7 phbC gene. Total DNA from A. doebereinerae (lane 1), A. brasilense Sp7 (lane 2), A. lipoferum (lane 3), A. irakense (lane 4), A. amazonense (lane 5), A. halopraeferens (lane 6), and E. coli (lane 7). The DNA was digested with EcoRI. Arrows indicate the estimated position of the putative phbC bands.

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.

Techniques: Southern Blot, Hybridization, Labeling, Generated

Journal:

Article Title: Identification and Isolation of Genes Involved in Poly(?-Hydroxybutyrate) Biosynthesis in Azospirillum brasilense and Characterization of a phbC Mutant

doi: 10.1128/AEM.68.6.2943-2949.2002

Figure Lengend Snippet: Growth curves of A. brasilense Sp7 (▴) and of the phbC mutant (•) in LB media. Bacteria were grown in a shaker (170 rpm at 30°C). Each value represents the mean of three replicates from one representative experiment. Each experiment was carried out three times, each yielding similar results.

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.

Techniques: Mutagenesis

Journal:

Article Title: Identification and Isolation of Genes Involved in Poly(?-Hydroxybutyrate) Biosynthesis in Azospirillum brasilense and Characterization of a phbC Mutant

doi: 10.1128/AEM.68.6.2943-2949.2002

Figure Lengend Snippet: Aggregation of A. brasilense Sp7 (A) and of the phbC mutant (B) after 24 h of growth in medium with a high C:N ratio. Bacteria were transferred to Petri dishes before the photograph was taken.

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.

Techniques: Mutagenesis

Journal:

Article Title: Identification and Isolation of Genes Involved in Poly(?-Hydroxybutyrate) Biosynthesis in Azospirillum brasilense and Characterization of a phbC Mutant

doi: 10.1128/AEM.68.6.2943-2949.2002

Figure Lengend Snippet: Electron micrographs of thin sections of A. brasilense (A) and of the phbC mutant (B), grown for 24 h in medium with a high C:N ratio. The arrow indicates PHB granules. Bars, 1 μm. At least 10 sections of each strain were examined.

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.

Techniques: Mutagenesis

Journal:

Article Title: Identification and Isolation of Genes Involved in Poly(?-Hydroxybutyrate) Biosynthesis in Azospirillum brasilense and Characterization of a phbC Mutant

doi: 10.1128/AEM.68.6.2943-2949.2002

Figure Lengend Snippet: The effect of PHB on survival capability of starved bacteria. Cells of A. brasilense Sp7 (▴) and phbC mutant (•) were grown on medium with a high C:N ratio for 24 h and transferred to phosphate buffer, where they were incubated for 12 days. Bacterial density was determined using dilution plating. Each value represents the mean of three replicates from one representative experiment. Each experiment was carried out three times, each yielding similar results.

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.

Techniques: Mutagenesis, Incubation

Journal:

Article Title: Identification and Isolation of Genes Involved in Poly(?-Hydroxybutyrate) Biosynthesis in Azospirillum brasilense and Characterization of a phbC Mutant

doi: 10.1128/AEM.68.6.2943-2949.2002

Figure Lengend Snippet: Adhesion of A. brasilense Sp7 and phbC mutant to roots of sweet corn and wheat a

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.

Techniques: Mutagenesis

Journal:

Article Title: Identification and Isolation of Genes Involved in Poly(?-Hydroxybutyrate) Biosynthesis in Azospirillum brasilense and Characterization of a phbC Mutant

doi: 10.1128/AEM.68.6.2943-2949.2002

Figure Lengend Snippet: Extraction of extracellular polysaccharides from A. brasilense Sp7 and phbC mutant after 48 h of growth in high C:N-ratio medium a

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.

Techniques: Mutagenesis