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

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

Abstract

Pyrococcus and Thermococcus are hyperthermophilic archaea in the order Thermococcales. Bothgenera are strictly anaerobes, gaining energy by fermentation of peptide and several carbohydrates at optimaltemperature above 70 °C. A Pyrococcus sp. strain Pikanate 5017 (PK 5017) was recently isolated from ahot spring in Northern Thailand. The strain PK 5017 is a valuable source of numerous genes encodingthermostable enzymes. Growth kinetics determined at various temperatures (75-105 °C) indicates that strainPK5017 is a fast growing archaeon. An arbitrarily primed PCR (AP-PCR) technique was successfully appliedto 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 effectivein producing polymorphisms of the PCR products at 3-10 ng of DNAs. The unique AP-PCR fingerprintsdistinguish the strain PK5017 from P. furiosus, P. horikoshii, P. abyssi, T. litoralis, and T. celer. The resultsindicate a clear distinction of genome sequences among Pyrococcus and Thermococcus genera.

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How to Cite
Pasomsup, P., González, J. M., Portillo, M. C., Pongsapukdee, V., & Kanoksilapatham, W. (2013). Differentiation of a Hyperthermophilic Archaeon Pyrococcus sp. strain Pikanate 5017, by Arbitrarily Primed PCR. Science, Engineering and Health Studies, 5(1), 14–23. https://doi.org/10.14456/sustj.2011.10
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Research Articles

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