Characterization of the Replicon from Plasmid pAC1 from Acetobacter pasteurianus

Grones, Jozef; Králová, Alena; Turňa, Ján

doi:10.1006/bbrc.1993.1179

Cited by 9 publications

(10 citation statements)

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“…YPG (GRONES et al, 1989) and LB media were used for routine cultivation of both Acetobacter pasteurianus and Escherichia coli. When necessary, kanamycin (Km) and ampicillin (Ap) were added to the medium at a concentration of 30 µg/mL and 100 µg/mL, respectively.…”

Section: Methodsmentioning

confidence: 99%

“…Plasmid DNA from E. coli cells was isolated according to BIRNBOIM and DOLY (1979) and from A. pasteurianus cells using the modified method of GRONES et al (1989). Plasmids from sequence analyses were purified using the Wizard Plus Midipreps DNA Purification System (Promega).…”

Section: Isolation Of Plasmid Dnamentioning

confidence: 99%

“…Total DNA was isolated from the same amount of the cells (OD590 = 1.0) of each culture using the SDS-lysozyme lysis method (GRONES et al, 1989). Total DNA thus obtained was then purified by ethanol precipitation and examined using 1% agarose gel electrophoresis.…”

Section: Sequencing Of Pap4mentioning

confidence: 99%

“…Based on cryptic plasmid pAC1 from Acetobacter pasteurianus several plasmid vectors have been construed (Grones et al, 1991;Grones & Turňa, 1992). There vectors were used for cloning, successfully transformed and expressed not only in E. coli, Acetobacter, but they also were able to replicate in some Gram-negative and Gram-positive bacteria (Grones et al, 1993;Grones & Turňa, 1995).…”

Section: Introductionmentioning

confidence: 99%

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Molecular analysis of plasmid pAP4 from Acetobacter pasteurianus

Grones

Kretova

2006

Biologia

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Abstract:The complete nucleotide sequence of plasmid pAP4 isolated from Acetobacter pasteurianus 2374T has been determined. Plasmid pAP4 was analysed and found to be 3,870 bp in size with a G+C content of 50.1%. Computer assisted analysis of sequence data revealed 2 possible ORFs with typical promoter regions. ORF1 codes for a protein responsible for kanamycin resistance similar with Tn5 transposone, ORF2 encodes a resistance to ampicillin identical with Tn3 transposone. Plasmid has in A. pasteurianus five copies and in E. coli DH1 about 30 copies per chromosome and it segregation stability in both strains is very high. Based on the data on replication region, plasmid does not code for a replication protein and origin region is similar with ColE1-like plasmid.

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Section: Methodsmentioning

confidence: 99%

Section: Isolation Of Plasmid Dnamentioning

confidence: 99%

Section: Sequencing Of Pap4mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Molecular analysis of plasmid pAP4 from Acetobacter pasteurianus

Grones

Kretova

2006

Biologia

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“…The first identified cryptic plasmid from Acetobacter had been used for the construction of cloning vectors (Okumura et al 1985;Fukaya et al 1990;Tamaki et al 1991;Fujiwara et al 1992). Later a large plasmid pAC1 (19 kb) (Grones et al 1989;Grones & Turňa 1992), plasmid pAP12875 (Fomenkov et al 1995) and a large series of pAP1-pAP4 plasmids (Krahulec et al 2003) were indented from Acetobacter pasteurianus. Plasmid pJK2-1 (Trček et al 2000) and plasmid pAG20 (Kretová et al 2005) were isolated from Gluconobacter and Acetobacter aceti, respectively.…”

Section: Introductionmentioning

confidence: 99%

Nucleotide sequence analysis of small cryptic plasmid pGP2 from Acetobacter estunensis

Grones

2011

Biologia

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Complete nucleotide sequence of plasmid pGP2 from Acetobacter estunensis GP2 was identified after initial cloning of EcoRI fragment followed by preparation of deletion derivatives. Its size was defined to 2,797 bp and several sites for several restriction enzymes were revealed by DNA sequencing. Sequence analysis predicts three putative open reading frames (ORFs). ORF1 shows significant identity with the bacterial excinuclease α-subunit, ORF2 is a putative replication protein with low similarity with other Acetobacter plasmid's replication proteins, and ORF3 encodes a class B acid phosphatase/phosphotransferase. The replication module comprises a DnaA box like sequence, direct repeats, a potential prokaryotic promoter and a rep gene. The rep module is similar with several θ-replicating, iteron-containing modules from plasmids, suggesting pGP2 replication may follow the same course. Any phenotypic character determinant gene is absent in pGP2, suggesting this plasmid to be cryptic. However, a pGP2 derivative plasmid, containing the putative pGP2 rep region, can replicate and is stably maintained in Acetobacter and Escherichia coli strains; it can also carry foreign DNA fragments. Thus, pGP2-X could serve as a cloning shuttle vector between these bacteria. Prepared deletion derivatives of plasmid pGP2 suggested that Rep protein is essential for plasmid replication in host bacteria. In its natural host, A. estunensis GP2, pGP2 maintains a four-times lower copy number than in E. coli.

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Genetic organization ofAcetobacter for acetic acid fermentation

Beppu

1994

Antonie van Leeuwenhoek

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Plasmid vectors for the acetic acid-producing strains of Acetobacter and Gluconobacter were constructed from their cryptic plasmids and the efficient transformation conditions were established. The systems allowed to reveal the genetic background of the strains used in the acetic acid fermentation. Genes encoding indispensable components in the acetic acid fermentation, such as alcohol dehydrogenase, aldehyde dehydrogenase and terminal oxidase, were cloned and characterized. Spontaneous mutations at high frequencies in the acetic acid bacteria to cause the deficiency in ethanol oxidation were analyzed. A new insertion sequence element, IS1380, was identified as a major factor of the genetic instability, which causes insertional inactivation of the gene encoding cytochrome c, an essential component of the functional alcohol dehydrogenase complex. Several genes including the citrate synthase gene of A. aceti were identified to confer acetic acid resistance, and the histidinolphosphate aminotransferase gene was cloned as a multicopy suppressor of an ethanol sensitive mutant. Improvement of the acetic acid productivity of an A. aceti strain was achieved through amplification of the aldehyde dehydrogenase gene with a multicopy vector. In addition, spheroplast fusion of the Acetobacter strains was developed and applied to improve their properties.

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Characterization of the Replicon from Plasmid pAC1 from Acetobacter pasteurianus

Cited by 9 publications

References 0 publications

Molecular analysis of plasmid pAP4 from Acetobacter pasteurianus

Molecular analysis of plasmid pAP4 from Acetobacter pasteurianus

Nucleotide sequence analysis of small cryptic plasmid pGP2 from Acetobacter estunensis

Genetic organization ofAcetobacter for acetic acid fermentation

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