Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1987 Oct;169(10):4865–4868. doi: 10.1128/jb.169.10.4865-4868.1987

Cloning of plasmid genes encoding resistance to cadmium, zinc, and cobalt in Alcaligenes eutrophus CH34.

D Nies 1, M Mergeay 1, B Friedrich 1, H G Schlegel 1
PMCID: PMC213872  PMID: 2820947

Abstract

A 238-kilobase-pair plasmid, pMOL30, confers resistance to cadmium, zinc, and cobalt salts in Alcaligenes eutrophus CH34. After Tn5 mutagenesis, restriction nuclease analysis, and Southern DNA-DNA hybridization, a 9.1-kilobase-pair EcoRI fragment was found to harbor all of these resistance properties and was cloned into the broad-host-range hybrid plasmid pRK290. When transferred to a plasmid-free derivative of CH34, the hybrid plasmid conferred the same degree of resistance as the parent plasmid pMOL30. In two other Alcaligenes strains, the hybrid plasmid was expressed, but to a lower degree than in CH34 derivatives.

Full text

PDF
4865

Images in this article

4867Image
on p.4867

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bolivar F., Backman K. Plasmids of Escherichia coli as cloning vectors. Methods Enzymol. 1979;68:245–267. doi: 10.1016/0076-6879(79)68018-7. [DOI] [PubMed] [Google Scholar]
  2. Ditta G., Stanfield S., Corbin D., Helinski D. R. Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7347–7351. doi: 10.1073/pnas.77.12.7347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Foster T. J., Nakahara H., Weiss A. A., Silver S. Transposon A-generated mutations in the mercuric resistance genes of plasmid R100-1. J Bacteriol. 1979 Oct;140(1):167–181. doi: 10.1128/jb.140.1.167-181.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hansen J. B., Olsen R. H. Isolation of large bacterial plasmids and characterization of the P2 incompatibility group plasmids pMG1 and pMG5. J Bacteriol. 1978 Jul;135(1):227–238. doi: 10.1128/jb.135.1.227-238.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. KIHO Y., WATANABE I. The role of a trypsin-sensitive factor in multiplicity reactivation with heat-disrupted phage. J Mol Biol. 1961 Apr;3:125–132. doi: 10.1016/s0022-2836(61)80039-9. [DOI] [PubMed] [Google Scholar]
  6. Kado C. I., Liu S. T. Rapid procedure for detection and isolation of large and small plasmids. J Bacteriol. 1981 Mar;145(3):1365–1373. doi: 10.1128/jb.145.3.1365-1373.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Lejeune P., Mergeay M., Van Gijsegem F., Faelen M., Gerits J., Toussaint A. Chromosome transfer and R-prime plasmid formation mediated by plasmid pULB113 (RP4::mini-Mu) in Alcaligenes eutrophus CH34 and Pseudomonas fluorescens 6.2. J Bacteriol. 1983 Sep;155(3):1015–1026. doi: 10.1128/jb.155.3.1015-1026.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Mandel M., Higa A. Calcium-dependent bacteriophage DNA infection. J Mol Biol. 1970 Oct 14;53(1):159–162. doi: 10.1016/0022-2836(70)90051-3. [DOI] [PubMed] [Google Scholar]
  9. Mergeay M., Houba C., Gerits J. Extrachromosomal inheritance controlling resistance to cadmium, cobalt, copper and zinc ions: evidence from curing in a Pseudomonas [proceedings]. Arch Int Physiol Biochim. 1978 May;86(2):440–442. [PubMed] [Google Scholar]
  10. Mergeay M., Nies D., Schlegel H. G., 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. doi: 10.1128/jb.162.1.328-334.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Nakahara H., Silver S., Miki T., Rownd R. H. Hypersensitivity to Hg2+ and hyperbinding activity associated with cloned fragments of the mercurial resistance operon of plasmid NR1. J Bacteriol. 1979 Oct;140(1):161–166. doi: 10.1128/jb.140.1.161-166.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  13. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES