Biomics

Following the thermostable DNA polymerases from thermophilic eubacteria, attention was paid to microorganisms from another domain of Life, thermophilic and hyperthermophilic archaea, many of which are capable of growing at temperatures around 100 °С and above. Since 1991, more than three dozen genes of thermostable DNA polymerases of thermophilic and hyperthermophilic archaea representing 14 genera and about 30 species have been cloned and sequenced. DNA polymerases from the genus Thermococcus are the most widely represented. Pfu polymerase from the hyperthermophilic archaea Pyrococcus furiosus is the most commonly used. The vast majority of these enzymes belong to the B-family of DNA polymerases, and in addition to nucleotidyltransferase (polymerase) activity, they also have editing 3’→5’-exonuclease activity, which provides increased copy accuracy in PCR. Most of these archaeal DNA polymerases are characterized by a long half-life at 95 °С and even at 100 °С, measured for some by many hours. Archaeal DNA polymerases have found application in the amplification of difficult matrices, including those with long and high GC compositions, where mixtures of such enzymes with Taq polymerase have performed well.

archaea, hyperthermophilic archaea, DNA polymerase, thermostable DNA polymerase, 3’→5’-exonuclease editing activity, DNA, PCR

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