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Gene Synthesis> | … Construction of Recombinant Baculoviruses for rKOD. Thermococcus kodakarensis KOD DNA polymerase (Tkod-Pol, mature type, NCBI accession No. D 29671) gene was chemically synthesized (Genscript, Piscataway, USA) and cloned into pUC57 vector … | Get A Quote |
The KOD DNA polymerase from Thermococcus kodakarensis (Tkod-Pol) has been preferred for PCR due to its rapid elongation rate, extreme thermostability and outstanding fidelity. Here in this study, we utilized silkworm-baculovirus expression vector system (silkworm-BEVS) to express the recombinant Tkod-Pol (rKOD) with N-terminal (rKOD-N) or C-terminal (rKOD-C) tandem fusion tags. By using BEVS, we produced functional rKODs with satisfactory yields, about 1.1 mg/larva for rKOD-N and 0.25 mg/larva for rKOD-C, respectively. Interestingly, we found that rKOD-C shows higher thermostability at 95 °C than that of rKOD-N, while that rKOD-N is significantly unstable after exposing to long period of heat-... More