| 摘要 |
FIELD: genetic engineering, biochemistry, enzymology. SUBSTANCE: thermostable DNA polymerase is isolated and purified from archaebacterium Thermococcus gorgonarius that is able to retain enzymatic activity for 2 h in incubation at temperature 95 C completely practically in the presence of stabilizing agent. Enzyme shows high fidelity degree in replication. Also, DNA sequence encoding this new DNA polymerase is isolated and characterized. After expression of this nucleotide sequence in E. coli cells recombinant form of this enzyme is obtained. New DNA polymerase from Thermococcus gorgonarius can be used in methods for amplification and/or sequencing DNA where copying fidelity is provided being it exceeds this index by two-fold as compared with any of known DNA polymerases. Invention provides preparing native or recombinant form of thermostable DNA polymerase using in diagnostic and other investigations based on PCR (polymerase chain reaction)-amplification of nucleic acids. EFFECT: improved preparing method, valuable biological and biochemical properties of enzyme. 11 cl, 10 dwg, 1 tbl, 8 ex |
