Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Plasmid pHH1 of Halobacterium salinarium: characterization of the replicon region, the gas vesicle gene cluster and insertion elements

  • 89 Accesses

  • 51 Citations


The DNA sequence of the 5.7 kb plasmid pHH9 containing the replicon region of the 150 kb plasmid pHH1 from Halobacterium salinarium was determined. The minimal region necessary for stable plasmid maintenance lies within a 2.9 kb fragment, as defined by transformation experiments. The DNA sequence contained two open reading frames arranged in opposite orientations, separated by an unusually high AT-rich (60–70% A + T) sequence of 350 bp. All H. salinarium strains (H. halobium, H. cutirubrum) investigated harbour endogenous plasmids containing the pHH1 replicon; however, these pHH1-type plasmids differ by insertions and deletions. Adjacent to the replicon, and separated by a copy of each of the insertion elements ISH27 and ISH26, is the 9 kb p-vac region required for gas vesicle synthesis. Analysis of these and other ISH element copies in pHH1 revealed that most of them lack the target DNA duplication usually found with recently transposed ISH elements. These results underline the plasticity of plasmid pHH1.

This is a preview of subscription content, log in to check access.


  1. Blaseio U, Pfeifer F (1990) Transformation of Halobacterium halobium: Development of vectors and investigation of gas vesicle synthesis. Proc Natl Acad Sci USA 87:6772–6776

  2. Bramhill D, Kornberg A (1988) A model for initiation at origins of DNA replication. Cell 54:915–918

  3. Charlebois RL, Lam WL, Cline SW, Doolittle WF (1987) Characterization of pHV2 from Halobacterium volcanii and its use in demonstrating transformation of an archaebacterium. Proc Natl Acad Sci USA 84:8530–8534

  4. Charlebois RL, Schalkwyk L, Holman J, Doolittle WF (1991) Detailed physical map and set of overlapping cosmid clones covering the genome of the archaebacterium Haloferax volcanii DS2. Mol Biol 222:509–524

  5. Cline SW, Doolittle WF (1992) Transformation of members of the genus Haloarcula with shuttle vectors based on Halobacterium halobium and Haloferax volcanii plasmid replicons. J Bacteriol 174:1076–1080

  6. Cline SW, Schalkwyk L, Doolittle WF (1989) Transformation of the archaebacterium Halobacterium volcanii with genomic DNA. J Bacteriol 171:4987–4991

  7. DasSarma S (1989) Mechanisms of genetic variability in Halobacterium halobium: the purple membrane and gas vesicle mutations. Can J Microbiol 35:65–72

  8. DasSarma S, RajBhandari U, Khorana HG (1983) High-frequency spontaneous mutation in the bacterio-opsin gene in Halobacterium halobium is mediated by transposable elements. Proc Natl Acad Sci USA 80:2201–2205

  9. DasSarma S. Damerval T, Jones JG, Tandeau de Marsac N (1987) A plasmid-encoded gas vesicle protein gene in a halophilic archaebacterium. Mol Microbiol 1:365–370

  10. Devereux J, Häberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395

  11. Ebert K, Goebel W (1985) Conserved and variable regions in the chromosomal and extrachromosomal DNA of halobacteria. Mol Gen Genet 200:96–102

  12. Ebert K, Hanke C, Delius H, Goebel W, Pfeifer F (1987) A new insertion element, ISH26, from Halobacterium halobium. Mol Gen Genet 206:81–87

  13. Englert C, Krüger K, Offner S, Pfeifer F (1992) Three different but related gene clusters involved in gas vesicle synthesis in halophilic Archaea. J Mol Biol 227:586–592

  14. Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction fragments to high specific activity. Anal Biochem 132:6–13

  15. Hofman JD, Schalkwyk L, Doolittle WF (1986) ISH51: a large, degenerate family of insertion sequence-like elements in the genome of the archaebacterium Halobacterium volcanii. Nucleic Acids Res 14:6983–7000

  16. Holmes M, Nuttall S, Dyall-Smith M (1991) Construction and use of halobacterial shuttle vectors and further studies on Haloferax DNA gyrase. J Bacteriol 173:3807–3813

  17. Horne M, Englert C, Pfeifer F (1988) Two genes encoding gas vacuole proteins in Halobacterium halobium. Mol Gen Genet 213:459–464

  18. Horne M, Englert C, Wimmer C, Pfeifer F (1991) A DNA region of 9 kb contains all genes necessary for gas vesicle synthesis in halophilic archaebacteria. Mol Microbiol 5:1159–1174

  19. Jones JG, Hackett N, Halladay J, Scothorn D, Yang C, Ng W, DasSarma S (1989) Analysis of insertion mutants reveals two new genes in the gas vesicle gene cluster of Halobacterium halobium. Nucleic Acids Res 17:7785–7793

  20. Jones JG, Young DC, DasSarma S (1991) Structure and organization of the gas vesicle gene cluster on the Halobacterium halobium plasmid pNRC100. Gene 102:117–122

  21. Krebs M, RajBhandari U, Khorana HG (1990) Nucleotide sequence of ISH11, a new Halobacterium halobium insertion element isolated from the plasmid pGRB1. Nucleic Acids Res 18:6699

  22. Lam W, Doolittle WF (1989) Shuttle vectors for the archaebacterium Halobacterium volcanii. Proc Natl Acad Sci USA 86:5478–82

  23. Manen D, Caro L (1991) The replication of plasmid pSC101. Mol Microbiol 5:233–237

  24. Moore R, McCarthy B (1969) Characterization of the deoxyribonucleic acid of various strains of halophilic bacteria. J Bacteriol 99:248–254

  25. Ng WL, Kothakota S, DasSarma S (1991) Structure of the gas vesicle plasmid in Halobacterium halobium: inversion isomers, inverted repeats, and insertion sequences. J Bacteriol 173:1958–1964

  26. Pfeifer F (1985) Insertion elements and genome organization of Halobacterium halobium. Syst Appl Microbiol 7:36–40

  27. Pfeifer F (1988) Genetics of halobacteria. In: F Rodriguez-Valera (ed) Halophilic bacteria. Vol 2, CRC Press, Boca Raton Florida, pp 105–133

  28. Pfeifer F, Betlach M (1985) Genome organization in Halobacterium halobium: a 70 kb island of more AT-rich DNA in the chromosome. Mol Gen Genet 198:449–455

  29. Pfeifer F, Blaseio U (1989) Insertion elements and deletion formation in a halophilic archaebacterium. J Bacteriol 171:5135–5140

  30. Pfeifer F, Blaseio U (1990) Transposition burst of the ISH27 insertion element family in Halobacterium halobium. Nucleic Acids Res 18:6921–6952

  31. Pfeifer F, Ghahraman P (1991) The halobacterial insertion element ISH28. Nucleic Acids Res 19:5788

  32. Pfeifer F, Weidinger G, Goebel W (1981 a) Characterization of plasmids in halobacteria. J Bacteriol 145:369–374

  33. Pfeifer F, Weidinger G, Goebel W (1981 b) Genetic variability in Halobacterium halobium. J Bacteriol 145:375–381

  34. Pfeifer F, Ebert K, Weidinger G, Goebel W (1982) Structure and functions of chromosomal and extrachromosomal DNA in halobacteria. Zbl Bakt Hyg I Abt Orig. C3 110–119

  35. Pfeifer F, Betlach M, Martienssen R, Friedman J, Boyer HW (1983) Transposable elements of Halobacterium halobium. Mol Gen Genet 191:182–188

  36. Pfeifer F, Boyer HW, Betlach M (1985) Restoration of bacterioopsin gene expression in a revertant of Halobacterium halobium. J Bacteriol. 164:414–420

  37. Pfeifer F, Blaseio U, Ghahraman P (1988) Dynamic plasmid populations in Halobacterium halobium. J Bacteriol 170:3718–42

  38. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

  39. Simon RD (1978) Halobacterium strain 5 contains a plasmid which is correlated with the presence of gas vacuoles. Nature, London 273:314–317

  40. Simsek M, DasSarma S, RajBhandari U, Khorana HG (1981) A transposable element from Halobacterium halobium which inactivates the bacteriorhodopsin gene. Proc Natl Acad Sci USA 79:7268–7272

  41. Staley J, Marvin P. Pfennig N, Holt J (1989) Bergey's manual of systematic bacteriology. Vol. 3, Williams and Wilkins, Baltimore, Maryland, pp 2216–2228

  42. Weidinger G, Klotz G, Goebel W (1979) A large plasmid from Halobacterium halobium carrying genetic information for gas vacuole formation. Plasmid 2:377–386

  43. Weidinger G, Pfeifer F, Goebel W (1981) Plasmids in halobacteria: restriction maps. Methods Enzymol 88:374–379

  44. Xu WL, Doolittle WF (1983) Structure of the archaebacterial transposable element ISH50. Nucleic Acids Res 11:4195–4199

  45. Zvyaga TA, Zozulya SA, Guriev SO (1987) The nucleotide sequence of the archaebacterial transposable genetic element ISH S1. Bioorg Chem 13:1351–1357

Download references

Author information

Correspondence to Felicitas Pfeifer.

Additional information

Communicated by W. Goebel

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Pfeifer, F., Ghahraman, P. Plasmid pHH1 of Halobacterium salinarium: characterization of the replicon region, the gas vesicle gene cluster and insertion elements. Molec. Gen. Genet. 238, 193–200 (1993).

Download citation

Key words

  • Halobacterial transformation
  • Transposable elements
  • Deletions
  • Inversions
  • Rearrangements