Characterization and Replication Properties of the Zymomonas mobilis ATCC 10988 Plasmids pZMO1 and pZMO2

Arvanitis, Nikolaos; Pappas, Katherine-Maria; Kolios, George; Afendra, Amalia S.; Typas, Milton A.; Drainas, Constantin

doi:10.1006/plas.2000.1480

Cited by 17 publications

(14 citation statements)

References 33 publications

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“…Four native plasmids from Z. mobilis ATCC 10988 have been used as the basis for the construction of shuttle vectors: pZMO1 (1,651 bp) [21], pZMO2 (1,669 bp) [20-22], pZM2 (pZMO3; 2,749 bp) [16,23,24] and pZM3 (pZMOB05; 4,023 bp) [17,25]. Shuttle vectors have also been constructed from native plasmids isolated from other Z. mobilis strains, such as pNSW301 from the ZM6100 strain [26]; pZMPI from the Z. mobilis PROM Al strain [27] and pZA2 from the NCIMB 8827 strain [19].…”

Section: Introductionmentioning

confidence: 99%

pZMO7-Derived shuttle vectors for heterologous protein expression and proteomic applications in the ethanol-producing bacterium Zymomonas mobilis

Chen

Lacap-Bugler

et al. 2014

BMC Microbiol

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BackgroundThe ethanol-producing bacterium Zymomonas mobilis has attracted considerable scientific and commercial interest due to its exceptional physiological properties. Shuttle vectors derived from native plasmids have previously been successfully used for heterologous gene expression in this bacterium for a variety of purposes, most notably for metabolic engineering applications.ResultsA quantitative PCR (qPCR) approach was used to determine the copy numbers of two endogenous double stranded DNA plasmids: pZMO1A (1,647 bp) and pZMO7 (pZA1003; 4,551 bp) within the NCIMB 11163 strain of Z. mobilis. Data indicated pZMO1A and pZMO7 were present at ca. 3-5 and ca. 1-2 copies per cell, respectively. A ca. 1,900 bp fragment from plasmid pZMO7 was used to construct two Escherichia coli – Z. mobilis shuttle vectors (pZ7C and pZ7-184). The intracellular stabilities and copy numbers of pZ7C and pZ7-184 were characterized within the NCIMB 11163, ATCC 29191 and (ATCC 10988-derived) CU1 Rif2 strains of Z. mobilis. Both shuttle vectors could be stably maintained within the ATCC 29191 strain (ca. 20-40 copies per cell), and the CU1 Rif2 strain (ca. 2-3 copies per cell), for more than 50 generations in the absence of an antibiotic selectable marker. A selectable marker was required for shuttle vector maintenance in the parental NCIMB 11163 strain; most probably due to competition for replication with the endogenous pZMO7 plasmid molecules. N-terminal glutathione S-transferase (GST)-fusions of four endogenous proteins, namely the acyl-carrier protein (AcpP); 2-dehydro-3-deoxyphosphooctonate aldolase (KdsA); DNA polymerase III chi subunit (HolC); and the RNA chaperone protein Hfq; were successfully expressed from pZ7C-derived shuttle vectors, and their protein-protein binding interactions were analyzed in Z. mobilis ATCC 29191. Using this approach, proteins that co-purified with AcpP and KdsA were identified.ConclusionsWe show that a shuttle vector-based protein affinity ‘pull-down’ approach can be used to probe protein interaction networks in Z. mobilis cells. Our results demonstrate that protein expression plasmids derived from pZMO7 have significant potential for use in future biological or biotechnological applications within Z. mobilis.

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Section: Introductionmentioning

confidence: 99%

pZMO7-Derived shuttle vectors for heterologous protein expression and proteomic applications in the ethanol-producing bacterium Zymomonas mobilis

Chen

Lacap-Bugler

et al. 2014

BMC Microbiol

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“…Bacterial conjugations were performed by triparental filter matings as previously described (Arvanitis et al, 2000). For counter selection against donor cells, 100mg/ml nalidixic acid was used for ATCC 10988.…”

Section: Bacterial Conjugationsmentioning

confidence: 99%

Differential expression of Zymomonas mobilis sucrase genes (sacB and sacC) in Escherichia coli and sucrase mutants of Zymomonas mobilis

Gurunathan

Gunasekaran

2004

Braz. arch. biol. technol.

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“…Among isolates deriving from various parts of the world, ATCC 10988, identified in the 1920s in Mexican pulque fermentations (initially called "Thermobacterium mobile") (17), is one of the longest-studied Zymomonas strains and has served for most biochemical and physiological works on the organism. ATCC 10988 exhibits fermenting performances that equal that of industrial strain CP4 in sucrose-rich media (15) and has also attracted attention due to its split genome: it bears multiple different plasmids, six of which are reported in this work, while the smallest have been cloned and sequenced in earlier works (2,10,12). Most species-specific vectors constructed and used for Z. mobilis, indeed, derive from the small multicopy plasmids of ATCC 10988 (16,19,21).…”

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confidence: 91%

Genome Sequence of the Ethanol-Producing Zymomonas mobilis subsp. mobilis Lectotype Strain ATCC 10988

et al. 2011

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Zymomonas mobilis ATCC 10988 is the type strain of the Z. mobilis subsp. mobilis taxon, members of which are some of the most rigorous ethanol-producing bacteria. Isolated from Agave cactus fermentations in Mexico, ATCC 10988 is one of the first Z. mobilis strains to be described and studied. Its robustness in sucrose-substrate fermentations, physiological characteristics, large number of plasmids, and overall genomic plasticity render this strain important to the study of the species. Here we report the finishing and annotation of the ATCC 10988 chromosomal and plasmid genome.

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Characterization and Replication Properties of the Zymomonas mobilis ATCC 10988 Plasmids pZMO1 and pZMO2

Cited by 17 publications

References 33 publications

pZMO7-Derived shuttle vectors for heterologous protein expression and proteomic applications in the ethanol-producing bacterium Zymomonas mobilis

pZMO7-Derived shuttle vectors for heterologous protein expression and proteomic applications in the ethanol-producing bacterium Zymomonas mobilis

Differential expression of Zymomonas mobilis sucrase genes (sacB and sacC) in Escherichia coli and sucrase mutants of Zymomonas mobilis

Genome Sequence of the Ethanol-Producing Zymomonas mobilis subsp. mobilis Lectotype Strain ATCC 10988

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