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DNA polymerases are present in all organisms and are important enzymes that synthesise DNA molecules. They are used in various fields of science, predominantly as essential components for  in vitro  DNA syntheses, known as PCR. Modern diagnostics, molecular biology and genetic engineering need DNA polymerases which demonstrate improved performance. This study was aimed at obtaining a new  Neq SSB -TaqS  fusion DNA polymerase from the  Taq  DNA Stoffel domain and a single-stranded DNA binding-like protein of  Nanoarchaeum equitans  in order to significantly improve the properties of DNA polymerase. The DNA coding sequence of  Taq  Stoffel DNA polymerase and the nonspecific DNA-binding protein of  Nanoarchaeum equitans  ( Neq SSB-like protein) were fused. A novel recombinant gene was obtained which was cloned into the pET-30 Ek/LIC vector and introduced into  E .  coli  for expression. The recombinant enzyme was purified and its enzymatic properties including DNA polymerase activity, PCR amplification rate, thermostability, processivity and resistance to inhibitors, were tested. The yield of the target protein reached approximately 18 mg/l after 24 h of the IPTG induction. The specific activity of the polymerase was 2200 U/mg. The recombinant  Neq SSB- Taq S exhibited a much higher extension rate (1000 bp template in 20 s), processivity (19 nt), thermostability (half-life 35 min at 95°C) and higher tolerance to PCR inhibitors (0.3–1.25% of whole blood, 0.84–13.5 μg of lactoferrin and 4.7–150 ng of heparin) than  Taq  Stoffel DNA polymerase. Furthermore, our studies show that  Neq SSB- Taq S DNA polymerase has a high level of flexibility in relation to Mg 2+  ions (from 1 to 5 mM) and KCl or (NH 4 ) 2 SO 4  salts (more than 60 mM and 40 mM, respectively). Using  Neq SSB- Taq S DNA polymerase instead of the  Taq  DNA polymerase could be a better choice in many PCR applications.

Fusion of Taq DNA polymerase with single-stranded DNA binding-like protein of Nanoarchaeum equitans—Expression and characterization