Abstract
This chapter provides methods and insights into the use of broad-host-range plasmid vectors useful for expression of genes in a variety of bacteria. The main focus is on IncQ, IncW, IncP, and pBBR1-based plasmids which have all been used for such applications. The specific design of each vector is adapted to its use, and here we describe, as an example, a protocol for construction (in Escherichia coli) of large insert DNA libraries in an IncP type vector and transfer of the library to the desired host. The genes of interest will in this case have to be expressed from their own promoters and the libraries will be screened by a method that best fits the functions of the gene or gene clusters searched for.
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References
Primrose S. B. and Twyman R. M. (2006) ‘Basic biology of plasmid and phage vectors’, Principles of gene manipulation (7th edn: Blackwell), 55–74.
del Solar G. and Espinosa M. (2000) Plasmid copy number control: an ever-growing story. Mol. Microbiol. 37, 492–500.
Datta N. (1979) ‘Plasmid classification: incompatibility grouping’, in K. Timmis and A. Puhler (eds.), Plasmids of Medical, Environmental and Commercial Importance (Amsterdam: Elsevier/North Holland), 3–12.
Novick R. P. (1987) Plasmid incompatibility. Microbiol. Rev. 51, 381–95.
Imanaka T. and Aiba S. (1981) A perspective on the application of genetic engineering: stability of recombinant plasmid. Ann. N.Y. Acad. Sci. 369, 1–14.
Aune T. E. V. and Aachmann F. L. (2010) Methodologies to increase the transformation efficiencies and the range of bacteria that can be transformed. Appl. Microbiol. Biotechnol. 85, 1301–13.
Figurski D. H. and Helinski D. R. (1979) Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc. Natl. Acad. Sci. U. S. A. 76, 1648–52.
Haring V. and Scherzinger E. (1989) ‘Replication proteins of the IncQ plasmid RSF1010’, in C. M. Thomas (ed.), Promiscuous Plasmids of Gram-Negative Bacteria (Academic Press, London, United Kingdom.), 95–124.
Fernández-López R., et al. (2006) Dynamics of the IncW genetic backbone imply general trends in conjugative plasmid evolution. FEMS Microbiol. Rev. 30, 942–66.
Valentine C. R. I. and Kado C. I. (1989) ‘Moelcular Genetics of IncW plasmids’, in C. M. Thomas (ed.), Promiscuous Plasmids of Gram-Negative Bacteria (Academic Press, London, United Kingdom.), 125–63.
Lefebre M. D. and Valvano M. A. (2002) Construction and evaluation of plasmid vectors optimized for constitutive and regulated gene expression in Burkholderia cepacia complex isolates. Appl. Environ. Microbiol. 68, 5956–64.
Vedler E., Vahter M., and Heinaru A. (2004) The completely sequenced plasmid pEST4011 contains a novel IncP1 backbone and a catabolic transposon harboring tfd genes for 2,4-dichlorophenoxyacetic acid degradation. J. Bacteriol. 186, 7161–74.
Schluter A., Szczepanowski R., Puhler A., and Top E. M. (2007) Genomics of IncP-1 antibiotic resistance plasmids isolated from wastewater treatment plants provides evidence for a widely accessible drug resistance gene pool. FEMS Microbiol. Rev. 31, 449–77.
Thomas C. M. and Helinski D. R. (1989) ‘Vegetative replication and stable inheritance of IncP plasmids’, in C. M. Thomas (ed.), Promiscuous Plasmids of Gram-Negative Bacteria (Academic Press, London, United Kingdom.), 1–25.
Bates S., Cashmore A. M., and Wilkins B. M. (1998) IncP plasmids are unusually effective in mediating conjugation of Escherichia coli and Saccharomyces cerevisiae: involvement of the tra2 mating system. J. Bacteriol. 180, 6538–43.
Poyart C. and Trieu-Cuot P. (1997) A broad-host-range mobilizable shuttle vector for the construction of transcriptional fusions to beta-galactosidase in gram-positive bacteria. FEMS Microbiol. Lett. 156, 193–98.
Waters V. L. (2001) Conjugation between bacterial and mammalian cells. Nat. Genet. 29, 375–76.
Burkardt H. J., Riess G., and Puhler A. (1979) Relationship of group P1 plasmids revealed by heteroduplex experiments: RP1, RP4, R68 and RK2 are identical. J. Gen. Microbiol. 114, 341–8.
Currier T. C. and Morgan M. K. (1981), Restriction endonuclease analyses of the incompatibility group P-1 plasmids RK2, RP1, RP4, R68, and R68.45. Curr. Microbiol. 5, 323–27.
Aakvik T., et al. (2009) A plasmid RK2-based broad-host-range cloning vector useful for transfer of metagenomic libraries to a variety of bacterial species. FEMS Microbiol. Lett. 296, 149–58.
Blatny J. M., Brautaset T., Winther-Larsen H. C., Karunakaran P., and Valla S. (1997) Improved broad-host-range RK2 vectors useful for high and low regulated gene expression levels in gram-negative bacteria. Plasmid 38, 35–51.
Sia E. A., Roberts R. C., Easter C., Helinski D. R., and Figurski D. H. (1995) Different relative importances of the par operons and the effect of conjugal transfer on the maintenance of intact promiscuous plasmid RK2. J. Bacteriol. 177, 2789–97.
Liles M. R., et al. (2009) Isolation and cloning of high-molecular-weight metagenomic DNA from soil microorganisms, Cold Spring Harb. Protoc. 2009, pdb.prot5271.
de Lorenzo V., Eltis L., Kessler B., and Timmis K. N. (1993) Analysis of Pseudomonas gene products using lacIq/Ptrp-lac plasmids and transposons that confer conditional phenotypes. Gene 123, 17–24.
Sambrook J. and Russel D. (2000) Molecular cloning: a laboratory manual (Cold Spring Harbor Laboratory Press, New York, N.Y.).
Bagdasarian M. M., Amann E., Lurz R., Rückert B., and Bagdasarian M. (1983) Activity of the hybrid trp-lac (tac) promoter of Escherichia coli in Pseudomonas putida. Construction of broad-host-range, controlled-expression vectors. Gene 26, 273–82.
Huang H. H., Camsund D., Lindblad P., and Heidorn T. (2010) Design and characterization of molecular tools for a Synthetic Biology approach towards developing cyanobacterial Âbiotechnology. Nucleic Acids Res. 38, 2577–93.
Schofield D. A., et al. (2003) Development of a thermally regulated broad-spectrum promoter system for use in pathogenic gram-positive Âspecies. Appl. Environ. Microbiol. 69, 3385–92.
Smits T. H., Seeger M. A., Witholt B., and van Beilen J. B. (2001) New alkane-responsive expression vectors for Escherichia coli and Pseudomonas. Plasmid 46, 16–24.
Newman J. R. and Fuqua C. (1999) Broad-host-range expression vectors that carry the L-arabinose-inducible Escherichia coli araBAD promoter and the araC regulator. Gene 227, 197–203.
Prior J. E., Lynch M. D., and Gill R. T. (2010) Broad-host-range vectors for protein expression across gram negative hosts. Biotechnol. Bioeng. 106, 326–32.
Sukchawalit R., Vattanaviboon P., Sallabhan R., and Mongkolsuk S. (1999). Construction and characterization of regulated L-arabinose-inducible broad host range expression vectors in Xanthomonas. FEMS Microbiol. Lett. 181, 217–23.
Singer J. T., et al. (2010) Broad-host-range plasmids for red fluorescent protein labeling of gram-negative bacteria for use in the zebrafish model system. Appl. Environ. Microbiol. 76, 3467–74.
Katzke N., et al. (2010) A novel T7 RNA polymerase dependent expression system for high-level protein production in the phototrophic bacterium Rhodobacter capsulatus. Protein Expression Purif. 69, 137–46.
Jeske M. and Altenbuchner J. (2010) The Escherichia coli rhamnose promoter rhaP(BAD) is in Pseudomonas putida KT2440 independent of Crp-cAMP activation. Appl. Microbiol. Biotechnol. 85, 1923–33.
Keil S. and Keil H. (1992) Construction of a cassette enabling regulated gene expression in the presence of aromatic hydrocarbons. Plasmid 27, 191–99.
Mermod N., Ramos J. L., Lehrbach P. R., and Timmis K. N. (1986) Vector for regulated expression of cloned genes in a wide range of gram-negative bacteria. J. Bacteriol. 167, 447–54.
Ramos J. L., Gonzäles-Carrero M., and Timmis K. N. (1988) Broad-host range expression vectors containing manipulated meta-cleavage pathway regulatory elements of the TOL plasmid. FEBS Lett. 226, 241.
Davison J., Heusterspreute M., Chevalier N., Ha-Thi V., and Brunel F. (1987) Vectors with restriction site banks. V. pJRD215, a wide-host-range cosmid vector with multiple cloning sites. Gene 51, 275–80.
Davison J. (2002) Genetic tools for pseudomonads, rhizobia, and other gram-Ânegative bacteria. BioTechniques 32, 386–8, 90, 92–4, passim.
Frey J. and Bagdasarian M. (1989) ‘The Moelcular Biology of IncQ plasmids’, in C. M. Thomas (ed.), Promiscuous Plasmids of Gram-Negative Bacteria (Academic Press, London, United Kingdom.), 79–94.
Labes M., Pühler A., and Simon R. (1990) A new family of RSF1010-derived expression and lac-fusion broad-host-range vectors for gram-negative bacteria. Gene 89, 37–46.
Leemans J., et al. (1982) Broad-host-range cloning vectors derived from the W-plasmid Sa. Gene 19, 361–64.
Antoine R. and Locht C. (1992) Isolation and molecular characterization of a novel broad-host-range plasmid from Bordetella bronchiseptica with sequence similarities to plasmids from gram-positive organisms. Mol. Microbiol. 6, 1785–99.
Kovach M. E., et al. (1995) Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes. Gene 166, 175–76.
Wild J., Hradecna Z., and Szybalski W. (2002) Conditionally amplifiable BACs: switching from single-copy to high-copy Âvectors and genomic clones. Genome Res. 12, 1434–44.
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Lale, R., Brautaset, T., Valla, S. (2011). Broad-Host-Range Plasmid Vectors for Gene Expression in Bacteria. In: Williams, J. (eds) Strain Engineering. Methods in Molecular Biology, vol 765. Humana Press. https://doi.org/10.1007/978-1-61779-197-0_19
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DOI: https://doi.org/10.1007/978-1-61779-197-0_19
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