Abstract
Over 60 Greenland glacial isolates were screened for plasmids and antibiotic resistance/sensitivity as the first step in establishing a genetic system. Sequence analysis of a small, cryptic, 1,950 bp plasmid, p54, from isolate GIC54, related to Arthrobacter agilis, showed a region similar to that found in theta replicating Rhodococcus plasmids. A 6,002 bp shuttle vector, pSVJ21, was constructed by ligating p54 and pUC18 and inserting a chloramphenicol acetyl transferase (CAT) cassette conferring chloramphenicol resistance. Candidate Gram-positive recipients were chosen among glacial isolates based on phylogenetic relatedness, relatively short doubling times at low temperatures, sensitivity to antibiotics, and absence of indigenous plasmids. We developed an electroporation protocol and transformed seven isolates related to members of the Arthrobacter, Microbacterium, Curtobacterium, and Rhodoglobus genera with pSVJ21. Plasmid stability was demonstrated by successive transformation into Escherichia coli and four Gram-positive isolates, growth without antibiotic, and plasmid re-isolation. This shuttle vector and our transformation protocol provide the basis for genetic experiments with different high G+C Gram-positive hosts to study cold adaptation and expression of cold-active enzymes at low temperatures.
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References
Alpert C, Crutz-Le Coq A, Malleret C, Zagoree M (2003) Characterization of a theta-type plasmid from Lactobacillus sakei: a potential basis for low-copy number vectors in lactobacilli. Appl Environ Microbiol 69:5574–5584
Bartoshik D, Baj J, Sochacka M, Piechucka E, Wlodarczuk M (2002) Molecular characterization of functional modules of plasmid pWKS1 of Paracoccus pantotrophus DSM 11072. Microbiology 148:2847–2856
Bauer AW, Kirby WM, Sherris JC, Turck M (1966) Antibiotic susceptibility testing by a standardized single disc method. Am J Clin Pathol 45:493–496
Brigidi P, De Rossi E, Bertarini ML, Riccardi G, Matteuzzi D (1990) Genetic transformation of Bacillus subtilis by electroporation. FEMS Microbiol Lett 67:135–138
Cavicchioli R, Ostrowski M (2003) Ultramicrobacteria. Encyclopedia of life sciences McMillan Publishers Ltd, Nature Publishing Group. http://www.els.net
Chattoraj DK (2000) Control of plasmid DNA replication by iterons: no longer paradoxical. Mol Microbiol 37(3):467–476
D’Costa VM, McGrann KM, Hughes DW, Wright GD (2006) Sampling the antibiotic resistome. Science 311:374–377
Cusano AM, Parrili E, Marino G, Tutino ML (2006) A novel genetic system for recombinant protein secretion in the Antarctic Pseudoalteromonas haloplanktis TAC125. Microb Cell Fact 5:40. doi:10.1186/1475-2859-5-40
del Solar G, Giraldo R, Ruiz-Echevarria MJ, Espinosa M, Diaz-Orejas R (1998) Replication and control of circular bacterial plasmids. Microbiol Mol Biol Rev 62:434–464
Desomer J, Dhaese P, van Mantagu M (1990) Transformation of Rhodococcus fascians by high-voltage electroporation and development of R. fascians cloning vectors. Appl Environ Microbiol 56:2818–2825
Hogan D, Kolter R (2002) Why are bacteria refractory to antimicrobials? Curr Opin Microbiol 5:472–477
Kobori H, Sullivan CW, Shizuya H (1984) Bacterial plasmids in Antarctic natural microbial assemblages. Appl Environ Microbiol 48(3):515–518
Kulakov LA, Larkin MJ, Kulakova AN (1997) Cryptic plasmid pKA22 isolated from the naphthalene degrading derivative of Rhodococcus rhodochrous NCIMB13064. Plasmid 38:61–69
Lessard PA, O’Brien XM, Currie DH, Sinskey AJ (2004) pB264, a small, mobilizable, temperature sensitive plasmid from Rhodococcus. BMC Microbiol 4(15):1–14
Ma Y, Wang L, Shao Z (2006) Pseudomonas, the dominant polycyclic aromatic hydrocarbon-degrading bacteria isolated from Antarctic soils and the role of large plasmids in horizontal gene transfer. Environ Microbiol 8(3):455–465
Miteva V, Sheridan P, Brenchley J (2004) Phylogenetic and physiological diversity of microorganisms isolated from a deep Greenland ice core. Appl Environ Microbiol 70:202–213
Miteva V, Brenchley J (2005) Detection and isolation of ultrasmall microorganisms from a 120,000 year old deep Greenland glacier ice core. Appl Environ Microbiol 71(12):7806–7818
Miyake R, Kawamoto J, Wei YL, Kitagawa M, Kato I, Kurihara T, Esaki N (2007) Construction of a low-temperature protein expression system using a cold adapted bacterium Shewanella sp. Strain Ac10, as a host. Appl Environ Microbiol 73(15):4849–4856
Morikawa M, Daido H, Pongpobpibool S, Imanaka T (1994) Construction of a new host-vector system in Arthrobacter sp. and cloning of the lipase gene. Appl Microbiol Biotechnol 42(2–3):300–303
Na KS, Nagayasu K, Kuroda A, Takiguchi N, Ikeda T, Ohtake H, Kato J (2005) Development of a genetic transformation system for benzene tolerant Rhodococcus opacus strains. J Biosci Bioeng 99(4):408–414
Nakashima N, Tamura T (2004) A novel system for expressing recombinant proteins over a wide temperature range from 4 to 35°C. Biotechnol Bioeng 86:136–148
Osborn M (1999) Database of plasmid replicons. University of Essex, Essex. http://www.essex.ac.uk/bs/staff/osborn/DPR_home.htm
Overhage J, Sieker S, Homburg S, Parschat K, Fetzener S (2005) Identification of large linear plasmids in Arthrobacter spp encoding the degradation of quinaldine to anthranilate. Microbiology 151:491–500
Papa R, Rippa V, Sannia G, Marino G, Duilio A (2007) A effective cold inducible expression system developed in Pseudoalteromonas haloplanktis TAC125. J Biotechnol 127:199–210
Ray MK, Seshu Kumar G, Shivaji S (1991) Plasmids from the soil bacteria of Schirmacher Oasis, Antarctica. Microbios 67:151–157
Sandu C, Chiribau CB, Sachelaru P, Brandsch R (2005) Plasmids for nicotine-dependent and independent gene expression in Arthrobacter nicotinovorans and other Arthrobacter species. Appl Environ Microbiol 71(12):8920–8924
Shareck J, Choi Y, Lee B, Miguez CB (2004) Cloning vectors based on cryptic plasmids isolated from lactic acid bacteria: their characteristics and potential applications in biotechnology. Crit Rev Biotechnol 24(4):155–208
Shaw PC, Hartley BC (1988) A host-vector system for an Arthrobacter species. J Gen Microbiol 134(4):903–911
Shen M, Fang P, Xu D, Zhang Y, Cao W, Zhu Y, Zhao J, Qin Z (2006) Replication and inheritance of Nocardia plasmid pC1. FEMS Microbiol Lett 261:47–52
Smith K, Youngman P (1992) Use of a new integrational vector to investigate compartment-specific expression of the Bacillus subtilis spoIIM gene. Biochimie 74:705–711
Tutino M, Duilio A, Moretti M, Sannia G, Marino G (2000) A rolling-circle plasmid from Psychrobacter sp. TA144: evidence for a novel Rep subfamily. Biochem Biophys Res Commun 274:488–495
Tutino ML, Duilio A, Parrilli E, Remaut E, Sannia G, Marino G (2001) A novel replication element from an Antarctic plasmid as a tool for the expression of proteins at low temperature. Extremophiles 5:257–264
Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119
Acknowledgments
We thank Michelle Stewart and Jayson Trusa for the technical help. This research was supported by NSF Grant MO 0347475 and DOE Grant DE-FG02-93ER20117. Sarah Lantz received support from the WISE program of Penn State.
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Communicated by K. Horikoshi.
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Miteva, V., Lantz, S. & Brenchley, J. Characterization of a cryptic plasmid from a Greenland ice core Arthrobacter isolate and construction of a shuttle vector that replicates in psychrophilic high G+C Gram-positive recipients. Extremophiles 12, 441–449 (2008). https://doi.org/10.1007/s00792-008-0149-7
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DOI: https://doi.org/10.1007/s00792-008-0149-7