استخراج و خالص سازي آنزيم DNA پلي مراز باسيلوس كالدوتناكس جدا شده از آب گرم قاينارجه سبلان

Extraction and Purifcation of DNA Polymerise Enzyme from Bacillus Caldotenax Isolated from Gainarjah Hot Spa in Sabalan

DNA polymerase enzyme undertakes the DNA replication and in molecular research , DNA polymerase enzymes , especially thermostable enzymes , have great importance . In this reseach , thermostable DNA polymerase is extracted from Bacillus caldotenax isolated from Gainarjah hot spa in Sabalan region . The Bacteria were cultured in Luria-Bertani broth and harvested in logarithm phase . Crude cell extracts were prepared using lysozyme and sonication . Purification was performed by ammonium sulfate precipitation and three steps of chromatography with DEAE-sephadex , phosphocellulose and Heparin sepharose . DNA polymerase enzyme was purified 124-fold and specific activity of the purified enzyme was 216.7 (U/mg). DNA polymerase had the highest activity at 70°c . The molecular weight of the enzyme was determined to be approximately 92000 dalton using SDS-PAGE .