SILPAKORN UNIVERSITY SCIENCE AND TECHNOLOGY JOURNAL, Vol 5 No 1 (January - June) 2011

Differentiation of a Hyperthermophilic Archaeon Pyrococcus sp. strain Pikanate 5017, by Arbitrarily Primed PCR

Patlada Pasomsup, Juan Miguel González, Maria Carmen Portillo, Veeranun Pongsapukdee, Wirojne Kanoksilapatham

Department of Microbiology, Faculty of Science, Silpakorn University, Nakhon Pathom

IRNAS-CSIC Avda. Reina Mercedes 10, 41012 Sevilla

IRNAS-CSIC Avda. Reina Mercedes 10, 41012 Sevilla

Department of Statistics, Faculty of Science, Silpakorn University, Nakhon Pathom

Department of Microbiology, Faculty of Science, Silpakorn University, Nakhon Pathom


Abstract


Pyrococcus 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.

Key Words: Thermococcales; Pyrococcus; Thermococcus; fingerprint; hyperthermophile; hot spring

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