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J Bacteriol. Sep 1989; 171(9): 5065–5070.
PMCID: PMC210318

Cloning and expression of plasmid genes encoding resistances to chromate and cobalt in Alcaligenes eutrophus.

Abstract

Resistances to chromate and cobalt were cloned on a 30-kilobase-pair (kb) DNA region from the large Alcaligenes eutrophus plasmid pMOL28 into the broad-host-range mobilizable cosmid vector pVK102. A restriction nuclease map of the 30-kb region was generated. The resistances expressed from the hybrid plasmids after transfer back into A. eutrophus were inducible and conferred the same degree of resistance as the parent plasmid pMOL28. Resistances were expressed in metal-sensitive Alcaligenes strains and related bacteria but not in Escherichia coli. Resistance to chromate was further localized on a 2.6-kb EcoRI fragment, and resistance to cobalt was localized on an adjoining 8.5-kb PstI-EcoRI fragment. When the 2.6-kb EcoRI fragment was expressed in E. coli under the control of a bacteriophage T7 promoter, three polypeptides with molecular masses of 31,500, 21,000, and 14,500 daltons were visible on autoradiograms. The 31,500- and 21,000-dalton polypeptides were membrane bound; the 14,500-dalton polypeptide was soluble.

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Selected References

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  • Don RH, Pemberton JM. Properties of six pesticide degradation plasmids isolated from Alcaligenes paradoxus and Alcaligenes eutrophus. J Bacteriol. 1981 Feb;145(2):681–686. [PMC free article] [PubMed]
  • Ames GF. Resolution of bacterial proteins by polyacrylamide gel electrophoresis on slabs. Membrane, soluble, and periplasmic fractions. J Biol Chem. 1974 Jan 25;249(2):634–644. [PubMed]
  • Hansen JB, Olsen RH. Isolation of large bacterial plasmids and characterization of the P2 incompatibility group plasmids pMG1 and pMG5. J Bacteriol. 1978 Jul;135(1):227–238. [PMC free article] [PubMed]
  • Holloway BW. Genetics of Pseudomonas. Bacteriol Rev. 1969 Sep;33(3):419–443. [PMC free article] [PubMed]
  • Knauf VC, Nester EW. Wide host range cloning vectors: a cosmid clone bank of an Agrobacterium Ti plasmid. Plasmid. 1982 Jul;8(1):45–54. [PubMed]
  • Mergeay M, Nies D, Schlegel HG, Gerits J, Charles P, Van Gijsegem F. Alcaligenes eutrophus CH34 is a facultative chemolithotroph with plasmid-bound resistance to heavy metals. J Bacteriol. 1985 Apr;162(1):328–334. [PMC free article] [PubMed]
  • Nies D, Mergeay M, Friedrich B, Schlegel HG. Cloning of plasmid genes encoding resistance to cadmium, zinc, and cobalt in Alcaligenes eutrophus CH34. J Bacteriol. 1987 Oct;169(10):4865–4868. [PMC free article] [PubMed]
  • Nies DH, Silver S. Plasmid-determined inducible efflux is responsible for resistance to cadmium, zinc, and cobalt in Alcaligenes eutrophus. J Bacteriol. 1989 Feb;171(2):896–900. [PMC free article] [PubMed]
  • Nies DH, Silver S. Metal ion uptake by a plasmid-free metal-sensitive Alcaligenes eutrophus strain. J Bacteriol. 1989 Jul;171(7):4073–4075. [PMC free article] [PubMed]
  • Ohtake H, Cervantes C, Silver S. Decreased chromate uptake in Pseudomonas fluorescens carrying a chromate resistance plasmid. J Bacteriol. 1987 Aug;169(8):3853–3856. [PMC free article] [PubMed]
  • Siddiqui RA, Benthin K, Schlegel HG. Cloning of pMOL28-encoded nickel resistance genes and expression of the genes in Alcaligenes eutrophus and Pseudomonas spp. J Bacteriol. 1989 Sep;171(9):5071–5078. [PMC free article] [PubMed]
  • Siddiqui RA, Schlegel HG, Meyer M. Inducible and constitutive expression of pMOL28-encoded nickel resistance in Alcaligenes eutrophus N9A. J Bacteriol. 1988 Sep;170(9):4188–4193. [PMC free article] [PubMed]
  • Tabor S, Richardson CC. A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1074–1078. [PMC free article] [PubMed]
  • Yanisch-Perron C, Vieira J, Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. [PubMed]

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