Skip to main content
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1985 Mar;49(3):686–692. doi: 10.1128/aem.49.3.686-692.1985

Preparation of a DNA gene probe for detection of mercury resistance genes in gram-negative bacterial communities.

T Barkay, D L Fouts, B H Olson
PMCID: PMC373572  PMID: 3994373

Abstract

A DNA gene probe was prepared to study genetic change mechanisms responsible for adaptation to mercury in natural bacterial communities. The probe was constructed from a 2.6-kilobase NcoI-EcoRI DNA restriction fragment which spans the majority of the mercury resistance operon (mer) in the R-factor R100. The range of specificity of this gene probe was defined by hybridization to the DNA of a wide variety of mercury-resistant bacteria previously shown to possess the mercuric reductase enzyme. All of the tested gram-negative bacteria had DNA sequences homologous to the mer probe, whereas no such homologies were detected in DNA of the gram-positive strains. Thus, the mer probe can be utilized to study gene flow processes in gram-negative bacterial communities.

Full text

PDF
686

Images in this article

Selected References

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

  1. Allen D. A., Austin B., Colwell R. R. Antibiotic resistance patterns of metal-tolerant bacteria isolated from an estuary. Antimicrob Agents Chemother. 1977 Oct;12(4):545–547. doi: 10.1128/aac.12.4.545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Altherr M. R., Kasweck K. L. In situ studies with membrane diffusion chambers of antibiotic resistance transfer in Escherichia coli. Appl Environ Microbiol. 1982 Oct;44(4):838–843. doi: 10.1128/aem.44.4.838-843.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barrineau P., Summers A. O. A second positive regulatory function in the mer (mercury resistance) operon. Gene. 1983 Nov;25(2-3):209–221. doi: 10.1016/0378-1119(83)90225-1. [DOI] [PubMed] [Google Scholar]
  4. Brown N. L., Ford S. J., Pridmore R. D., Fritzinger D. C. Nucleotide sequence of a gene from the Pseudomonas transposon Tn501 encoding mercuric reductase. Biochemistry. 1983 Aug 16;22(17):4089–4095. doi: 10.1021/bi00286a015. [DOI] [PubMed] [Google Scholar]
  5. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clark D. L., Weiss A. A., Silver S. Mercury and organomercurial resistances determined by plasmids in Pseudomonas. J Bacteriol. 1977 Oct;132(1):186–196. doi: 10.1128/jb.132.1.186-196.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Clewell D. B., Helinski D. R. Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form. Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159–1166. doi: 10.1073/pnas.62.4.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Crawford J. T., Bates J. H. Isolation of plasmids from mycobacteria. Infect Immun. 1979 Jun;24(3):979–981. doi: 10.1128/iai.24.3.979-981.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dagert M., Ehrlich S. D. Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells. Gene. 1979 May;6(1):23–28. doi: 10.1016/0378-1119(79)90082-9. [DOI] [PubMed] [Google Scholar]
  10. Echeverria P., Seriwatana J., Chityothin O., Chaicumpa W., Tirapat C. Detection of enterotoxigenic Escherichia coli in water by filter hybridization with three enterotoxin gene probes. J Clin Microbiol. 1982 Dec;16(6):1086–1090. doi: 10.1128/jcm.16.6.1086-1090.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fitts R., Diamond M., Hamilton C., Neri M. DNA-DNA hybridization assay for detection of Salmonella spp. in foods. Appl Environ Microbiol. 1983 Nov;46(5):1146–1151. doi: 10.1128/aem.46.5.1146-1151.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Forbes B. A., Schaberg D. R. Transfer of resistance plasmids from Staphylococcus epidermidis to Staphylococcus aureus: evidence for conjugative exchange of resistance. J Bacteriol. 1983 Feb;153(2):627–634. doi: 10.1128/jb.153.2.627-634.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Foster T. J. Plasmid-determined resistance to antimicrobial drugs and toxic metal ions in bacteria. Microbiol Rev. 1983 Sep;47(3):361–409. doi: 10.1128/mr.47.3.361-409.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fouts D. L., Manning J. E., Fox G. M., Schmid C. W. A complex repeated DNA sequence within the Drosophila transposable element copia. Nucleic Acids Res. 1981 Dec 21;9(24):7053–7064. doi: 10.1093/nar/9.24.7053. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fox B. S., Walsh C. T. Mercuric reductase: homology to glutathione reductase and lipoamide dehydrogenase. Iodoacetamide alkylation and sequence of the active site peptide. Biochemistry. 1983 Aug 16;22(17):4082–4088. doi: 10.1021/bi00286a014. [DOI] [PubMed] [Google Scholar]
  17. Graham J. P., Istock C. A. Gene exchange and natural selection cause Bacillus subtilis to evolve in soil culture. Science. 1979 May 11;204(4393):637–639. doi: 10.1126/science.107592. [DOI] [PubMed] [Google Scholar]
  18. Grunstein M., Hogness D. S. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. doi: 10.1073/pnas.72.10.3961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hill W. E., Payne W. L., Aulisio C. C. Detection and enumeration of virulent Yersinia enterocolitica in food by DNA colony hybridization. Appl Environ Microbiol. 1983 Sep;46(3):636–641. doi: 10.1128/aem.46.3.636-641.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
  21. Izaki K. Enzymatic reduction of mercurous and mercuric ions in Bacillus cereus. Can J Microbiol. 1981 Feb;27(2):192–197. doi: 10.1139/m81-030. [DOI] [PubMed] [Google Scholar]
  22. Jensen S., Jernelöv A. Biological methylation of mercury in aquatic organisms. Nature. 1969 Aug 16;223(5207):753–754. doi: 10.1038/223753a0. [DOI] [PubMed] [Google Scholar]
  23. Martin P. A., Dugan P. R., Tuovinen O. H. Plasmid DNA in acidophilic, chemolithotrophic thiobacilli. Can J Microbiol. 1981 Aug;27(8):850–853. doi: 10.1139/m81-133. [DOI] [PubMed] [Google Scholar]
  24. Meissner P. S., Falkinham J. O., 3rd Plasmid-encoded mercuric reductase in Mycobacterium scrofulaceum. J Bacteriol. 1984 Feb;157(2):669–672. doi: 10.1128/jb.157.2.669-672.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Miki T., Easton A. M., Rownd R. H. Mapping of the resistance genes of the R plasmid NR1. Mol Gen Genet. 1978 Jan 17;158(3):217–224. doi: 10.1007/BF00267192. [DOI] [PubMed] [Google Scholar]
  26. Morris C. F., Hashimoto H., Mickel S., Rownd R. Round of replication mutant of a drug resistance factor. J Bacteriol. 1974 Jun;118(3):855–866. doi: 10.1128/jb.118.3.855-866.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Morrison W. D., Miller R. V., Sayler G. S. Frequency of F116-mediated transduction of Pseudomonas aeruginosa in a freshwater environment. Appl Environ Microbiol. 1978 Nov;36(5):724–730. doi: 10.1128/aem.36.5.724-730.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. NAKAYA R., NAKAMURA A., MURATA Y. Resistance transfer agents in Shigella. Biochem Biophys Res Commun. 1960 Dec;3:654–659. doi: 10.1016/0006-291x(60)90081-4. [DOI] [PubMed] [Google Scholar]
  29. Nakahara H., Kinscherf T. G., Silver S., Miki T., Easton A. M., Rownd R. H. Gene copy number effects in the mer operon of plasmid NR1. J Bacteriol. 1979 Apr;138(1):284–287. doi: 10.1128/jb.138.1.284-287.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ni'Bhriain N. N., Silver S., Foster T. J. Tn5 insertion mutations in the mercuric ion resistance genes derived from plasmid R100. J Bacteriol. 1983 Aug;155(2):690–703. doi: 10.1128/jb.155.2.690-703.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Novick R. P., Roth C. Plasmid-linked resistance to inorganic salts in Staphylococcus aureus. J Bacteriol. 1968 Apr;95(4):1335–1342. doi: 10.1128/jb.95.4.1335-1342.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Nyman K., Nakamura K., Ohtsubo H., Ohtsubo E. Distribution of the insertion sequence IS1 in gram-negative bacteria. Nature. 1981 Feb 12;289(5798):609–612. doi: 10.1038/289609a0. [DOI] [PubMed] [Google Scholar]
  33. Ohtsubo H., Ohtsubo E. Nucleotide sequence of an insertion element, IS1. Proc Natl Acad Sci U S A. 1978 Feb;75(2):615–619. doi: 10.1073/pnas.75.2.615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Olson B. H., Barkay T., Colwell R. R. Role of plasmids in mercury transformation by bacteria isolated from the aquatic environment. Appl Environ Microbiol. 1979 Sep;38(3):478–485. doi: 10.1128/aem.38.3.478-485.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Olson G. J., Porter F. D., Rubinstein J., Silver S. Mercuric reductase enzyme from a mercury-volatilizing strain of Thiobacillus ferrooxidans. J Bacteriol. 1982 Sep;151(3):1230–1236. doi: 10.1128/jb.151.3.1230-1236.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Rawson M. V., Rogers W. A. Seasonal abundance of ancyrocephalinaen (Monogenoidea) parasites of bluegill, Lepomis macrochirus (RAF). J Wildl Dis. 1972 Jul;8(3):255–260. doi: 10.7589/0090-3558-8.3.255. [DOI] [PubMed] [Google Scholar]
  37. SILVERMAN M. P., LUNDGREN D. G. Studies on the chemoautotrophic iron bacterium Ferrobacillus ferrooxidans. I. An improved medium and a harvesting procedure for securing high cell yields. J Bacteriol. 1959 May;77(5):642–647. doi: 10.1128/jb.77.5.642-647.1959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Schottel J., Mandal A., Clark D., Silver S., Hedges R. W. Volatilisation of mercury and organomercurials determined by inducible R-factor systems in enteric bacteria. Nature. 1974 Sep 27;251(5473):335–337. doi: 10.1038/251335a0. [DOI] [PubMed] [Google Scholar]
  39. Silberstein Z., Cohen A. Hybridization analysis of restriction endonuclease DNA fragments of Bacillus cereus transcribed during spore outgrowth. J Bacteriol. 1978 Jun;134(3):1081–1088. doi: 10.1128/jb.134.3.1081-1088.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. So M., Heffron F., McCarthy B. J. The E. coli gene encoding heat stable toxin is a bacterial transposon flanked by inverted repeats of IS1. Nature. 1979 Feb 8;277(5696):453–456. doi: 10.1038/277453a0. [DOI] [PubMed] [Google Scholar]
  41. Spangler W. J., Spigarelli J. L., Rose J. M., Miller H. M. Methylmercury: bacterial degradation in lake sediments. Science. 1973 Apr 13;180(4082):192–193. doi: 10.1126/science.180.4082.192. [DOI] [PubMed] [Google Scholar]
  42. Stanisich V. A., Bennett P. M., Richmond M. H. Characterization of a translocation unit encoding resistance to mercuric ions that occurs on a nonconjugative plasmid in Pseudomonas aeruginosa. J Bacteriol. 1977 Mar;129(3):1227–1233. doi: 10.1128/jb.129.3.1227-1233.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Summers A. O., Kight-Olliff L. Tn1 generated mutants in the mercuric ion reductase of the Inc P plasmid, R702. Mol Gen Genet. 1980;180(1):91–97. doi: 10.1007/BF00267356. [DOI] [PubMed] [Google Scholar]
  44. Summers A. O., Lewis E. Volatilization of mercuric chloride by mercury-resistant plasmid-bearing strains of Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa. J Bacteriol. 1973 Feb;113(2):1070–1072. doi: 10.1128/jb.113.2.1070-1072.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Summers A. O., Silver S. Microbial transformations of metals. Annu Rev Microbiol. 1978;32:637–672. doi: 10.1146/annurev.mi.32.100178.003225. [DOI] [PubMed] [Google Scholar]
  46. Tanak N., Cramer J. H., Rownd R. H. EcoRI restriction endonuclease map of the composite R plasmid NR1. J Bacteriol. 1976 Jul;127(1):619–636. doi: 10.1128/jb.127.1.619-636.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Timoney J. F., Port J., Giles J., Spanier J. Heavy-metal and antibiotic resistance in the bacterial flora of sediments of New York Bight. Appl Environ Microbiol. 1978 Sep;36(3):465–472. doi: 10.1128/aem.36.3.465-472.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Tonomura K., Kanzaki F. The reductive decomposition of organic mercurials by cell-free extract of a mercury-resistant pseudomonad. Biochim Biophys Acta. 1969 Jun 17;184(1):227–229. doi: 10.1016/0304-4165(69)90124-x. [DOI] [PubMed] [Google Scholar]
  49. Weiss A. A., Murphy S. D., Silver S. Mercury and organomercurial resistances determined by plasmids in Staphylococcus aureus. J Bacteriol. 1977 Oct;132(1):197–208. doi: 10.1128/jb.132.1.197-208.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES