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Differentiation of a hyperthermophilic archaeon Pyrococcus sp. Strain Pikanate 5017, by arbitrarily primed PCR

AuthorsPasomsup, P.; González Grau, Juan Miguel ; Portillo Guisado, María del Carmen ; Pongsapukdee, V.; Kanoksilapatham, W.
Issue Date2011
PublisherSilpakorn University
CitationSilpakorn University Science and Technology Journal 5(1): 14-23 (2011)
AbstractPyrococcus and Thermococcus are hyperthermophilic archaea in the order Thermococcales. Both genera are strictly anaerobes, gaining energy by fermentation of peptide and several carbohydrates at optimal temperature above 70 ° C. A Pyrococcus sp. strain Pikanate 5017 (PK 5017) was recently isolated from a hot spring in Northern Thailand. The strain PK 5017 is a valuable source of numerous genes encoding thermostable enzymes. Growth kinetics determined at various temperatures (75-105 ° C) indicates that strain PK5017 is a fast growing archaeon. An arbitrarily primed PCR (AP-PCR) technique was successfully applied to differentiate the genome sequences of six members of Thermococcales. Two single primers, ARB-1f (5’ ATGAG GACT GAAA CCATT 3’) and ARB-2f (5’ GTAAA ACGA CGGC CAGT 3’), are effective in producing polymorphisms of the PCR products at 3-10 ng of DNAs. The unique AP-PCR fingerprints distinguish the strain PK5017 from P. furiosus, P. horikoshii, P. abyssi, T. litoralis, and T. celer. The results indicate a clear distinction of genome sequences among Pyrococcus and Thermococcus genera.
Identifiersissn: 1905-9159
Appears in Collections:(IRNAS) Artículos
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