| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | The DNA sequences of Phi29, PWT-WT, FWT-WT, PIPI-WT, FWT-MT, PWT-MT, FWT-D423A_S490R, FWT-D423A_S490A, FWT-D423A_F491A, FWT-S490A_F491A, and FWT-S490R_F491A with TEV cleavable His-tags at the C-terminus, were optimized toward Escherichia coli (E. coli) and synthesized by Genscript Biotechnology (Nanjing, China). These sequences were cloned into XbaI and XhoI sites of the pET21a plasmid. Restriction-endonucleases XbaI, XhoI and T4 DNA ligase were used for digestion and ligation (Genscript Biotechnology, Nanjing, China). | Get A Quote |
The properties of DNA polymerases isolated from thermophilic and mesophilic microorganisms, such as the thermophilic Geobacillus stearothermophilus (Bst) and mesophilic Bacillus subtilis phage (Phi29), have been widely researched. However, DNA polymerases in psychrophilic microorganisms remain poorly understood. In this study, we present for the first time the expression and functional characterization of DNA polymerases PWT-WT and FWT-WT from Psychrobacillus sp. BL-248-WT-3 and FJAT-21963. Enzymatic activity assays revealed that FWT-WT possessed strand displacement but lacked exonuclease activity and high ionic strength tolerance, whereas PWT-WT lacked all these properties. Further protein engineering and bioc... More
