Thermostable DNA-Polymerase from Thermus thermophilus B35: Isolation and Characterization of Some Properties

N. I. Rechkunova¹, A. G. Akishev², N. A. Lebedeva³, S. N. Khodyreva¹, S. Kh. Degtyarev^2*, and O. I. Lavrik¹

¹Novosibirsk Institute of Bioorganic Chemistry, Siberian Branch of the Russian Academy of Sciences, pr. Lavrent'eva 8, Novosibirsk, 630090 Russia; fax: (3832) 351-665

²Institute of Molecular Pathology and Ecological Biochemistry, Siberian Branch of the Russian Academy of Medical Sciences, ul. Timakova 2, Novosibirsk, 630117 Russia; fax: (3832) 336-853; E-mail: degt@ma.nsc.ru

³Novosibirsk State University, ul. Pirogova 2, Novosibirsk, 630090 Russia

^* To whom correspondence should be addressed.

Received January 5, 1998; Revision received June 4, 1998

---

Thermostable Tte DNA-polymerase was isolated from the strain Thermus thermophilus B35 which was found in hot spring water. The enzyme with molecular mass 87 kD was isolated using sequential chromatography on DEAE-Sepharose, hydroxylapatite, hexyl-agarose, and heparin-Sepharose. Biochemical properties of Tte DNA-polymerase are similar to those of Tth DNA-polymerase isolated from Thermus thermophilus HB8; however, practical application of Tte-Pol seems to be more favorable due to higher temperature optimum of this enzyme and lack of restriction endonucleases in the initial strain.

---

KEY WORDS: thermostable DNA-polymerase, isolation, properties, Thermus, DNA amplification

---