Analysis of replication region of the cryptic plasmid pAG20 from Acetobacter aceti 3620

Kretova, Miroslava; Szemes, Tomáš; Laco, Juraj; Gronesová, Paulína; Grones, Jozef

doi:10.1016/j.bbrc.2004.12.136

Cited by 5 publications

(11 citation statements)

References 17 publications

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“…The G+C content of the pGP2 plasmid DNA is 54.1% what is less than that for the A. estunensis chromosomal DNA (58.7%) (Cleenwerck et al 2002). Similar observation with higher G+C content of chromosomal DNA in comparison with the plasmid DNA was reported for small plasmids pAG20 from A. aceti CCM (Czech Collection of Microorganisms) 3610 (G+C 51.6%) (Kretová et al 2005) and pAP2 from A. pasteurianus CCM 2374 (G+C 51%) (Krahulec et al 2003).…”

Section: Resultssupporting

confidence: 78%

“…2). In the replication region a short repetitive sequence upstream of DnaA-binding sites was observed; similar to those described in plasmids pAG20 (Kretová et al 2005) and pJT2-1 (Trček et al 2000). Presence of a consensus sequence for the nic site within the ori region and repetitive sequences confirm that pGP2 plasmid is similar to ColE1 plasmids family.…”

Section: Analysis Of the Pgp2 Sequence And Putative Orfssupporting

confidence: 74%

“…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. One group of plasmids uses the Rep protein for initiation of replication, whereas the other group of plasmids (pAP1-pAP4) uses an RNA primer for initiation of replication (ColE1 like plasmids).…”

Section: Introductionmentioning

confidence: 99%

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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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Section: Resultssupporting

confidence: 78%

Section: Analysis Of the Pgp2 Sequence And Putative Orfssupporting

confidence: 74%

Section: Introductionmentioning

confidence: 99%

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Nucleotide sequence analysis of small cryptic plasmid pGP2 from Acetobacter estunensis

Grones

2011

Biologia

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“…Computer analysis of nucleotide sequence of plasmid pAP4 revealed eight putative ORFs longer than 200 nt. Only two ORFs, designated ORF1 and ORF2, were found to contain putative ribosome binding sites and promoter-like sequence similar to the consensus sequence of Acetobacter (Kretová et al, 2005), in their 5' upstream regions (Fig. 1).…”

Section: Resultsmentioning

confidence: 99%

“…pAH4 (Tonouchi et al 1994), pAP12875 (Fomenkov et al, 1995), pAP1 and pAP2 were isolated from Acetobacter pasteurianus (Krahulec et al, 2003), pJK2-1 from Acetobacter europaeus JK2 (Trček et al, 2000) and pAG20 from Acetobacter aceti 3620 (Kretová et al, 2005). Based on cryptic plasmid pAC1 from Acetobacter pasteurianus several plasmid vectors have been construed (Grones et al, 1991;Grones & Turňa, 1992).…”

Section: Introductionmentioning

confidence: 99%

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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The Rep20 Replication Initiator From the pAG20 Plasmid of Acetobacter aceti

et al. 2013

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In the previously isolated pAG20 plasmid from the Acetobacter aceti CCM3610 strain, the Rep20 protein was characterized as a main replication initiator. The pAG20 plasmid origin was localized in the vicinity of the rep20 gene and contained two 21-nucleotide-long iteron sequences, two 13-nucleotide-long direct repeats, and a DnaA-binding site. Electrophoretic mobility shift assay and nonradioactive fragment analysis confirmed that the Rep20 protein interacted with two direct repeats (5'-TCCAAATTTGGAT'-3') and their requirement during plasmid replication was verified by mutagenesis. Although the association could not be validated of the DnaA protein of from the host cells of Escherichia coli with the plasmid-encoded replication initiator that usually occurs during replication initiation, Rep20 was able to form dimeric structures by which it could bind the sequence of the rep20 gene and autoregulate its own expression. Targeted mutagenesis of the Rep20 protein revealed the importance of the third α-helix and ⁶³Lys, specifically during DNA binding. The second, closely adjacent β-sheet also took part in this process in which ⁵²Asn played a significant role.

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Analysis of replication region of the cryptic plasmid pAG20 from Acetobacter aceti 3620

Cited by 5 publications

References 17 publications

Nucleotide sequence analysis of small cryptic plasmid pGP2 from Acetobacter estunensis

Nucleotide sequence analysis of small cryptic plasmid pGP2 from Acetobacter estunensis

Molecular analysis of plasmid pAP4 from Acetobacter pasteurianus

The Rep20 Replication Initiator From the pAG20 Plasmid of Acetobacter aceti

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