Isolation of new plasmids from hyperthermophilic Archaea of the order Thermococcales

• Published in: Research in microbiology Vol. 148; no. 9; pp. 767 - 775
• Main Authors: Benbouzid-Rollet, N., López-García, P., Watrin, L., Erauso, G., Prieur, D., Forterre, P.
• Format: Journal Article
• Language: English
• Published: Paris Elsevier SAS 01.12.1997; Elsevier
• Subjects: Archaea; Bacteriology; Biological and medical sciences; Blotting, Southern; DNA, Bacterial - isolation & purification; Electrophoresis, Gel, Pulsed-Field; Fundamental and applied biological sciences. Psychology; Genetics; Hot Temperature; Hyperthermophile; Microbiology; Pacific Ocean; Plasmid; Plasmide; Plasmids - isolation & purification; Polymorphism, Restriction Fragment Length; Pyrococcus; Pyrococcus - chemistry; Pyrococcus - genetics; Restriction Mapping; Seawater; Thermococcales; Thermococcus; Thermococcus - chemistry; Thermococcus - genetics; Pacific Ocean; Plasmid; Thermococcales; Hyperthermophile; Archaea; Plasmide; Thermococcus; Pyrococcus; Genome organization; Archaeobacteria; Hyperthermophily; DNA; Bacteria; Thermococcaceae
• ISSN: 0923-2508; 1769-7123
• DOI: 10.1016/S0923-2508(97)82452-7

A collection of 57 strains of hyperthermophilic Archaea from the order Thermococcales was screened for the presence of plasmids; 9 plasmids present in six of these strains were isolated and characterized in terms of size and cross-hybridization. The Notl macrorestriction patterns of genomic DNA of strains harbouring these plasmids were obtained. Pyrococcus abyssi strains GE27 and GE23 as well as Thermococcus sp. GE31 contained a single plasmid of 3.5 kb (pGN27), 16.8 kb (pGN23) and 5.3 kb (pGN31), respectively, whilst the three strains 1559, 1560 and 1690 all contained two plasmids of 3.5 kb (pSN559, PSIM560, pSN690) and 24 kb (pLN559, pLN560, pLIM690), respectively. Plasmid pGN27 strongly cross-hybridized with the previously described plasmid pGT5 from P. abyssi strain GE5, whilst plasmids pGN23 and pGN31 did not cross-hybridize with each other, nor with any other plasmid. The three small plasmids of strains I559,1560 and I690 crosshybridized, as well as their three large plasmids. Macrorestriction pattern analysis and the results of plasmid cross-hybridization experiments indicated that these three strains were different but closely related, and likely belonged to the genus Thermococcus. This study shows that plasmids are widespread in hyperthermophilic archaea, and significantly increases the number and diversity of plasmids available for laboratory work. Une collection de 57 souches d'Archaea hyperthermophiles de l'ordre des Thermococcales a été criblée pour la présence de plasmides. Neuf plasmides, présents dans six de ces souches, ont été isolés et caractérisés. Les profils de macrorestriction NotI de l'ADN génomique des souches possédant ces plasmides sont présentés. Les souches de Pyrococcus abyssi GE27 et GE23, de même que la souche Thermococcus sp. GE31, possèdent un seul plasmide dont les tailles respectives sont de 3,5 kb (pGN27), 16,8 kb (pGN23) et 5,3 kb (pGN31). Le plasmide pGN27 hybride fortement avec le plasmide pGT5 de la souche GE5 de P. abyssi, tandis que les plasmides pGN23 et pGN31 ne donnent aucun signal en hybridation croisée, ni entre eux ni avec aucun des autres plasmides analysés. Les trois petits plasmides présents dans les souches 1559, 1560 et 1690 faybrident entre eux, ainsi que les trois grands plasmides présents dans ces mêmes souches. Le profil de macrorestriction et les résultats des expériences d'hybridation croisées montrent que ces trois souches sont différentes mais proches, et appartiennent probablement au genre Thermococcus. Ce travail montre que les plasmides sont répandus chez les Archaea hyperthermophiles et augmente de façon significative le nombre et la diversité des plasmides disponibles pour l'étude de ces microorganismes.