Novel properties of recombinant Sso7d-Taq DNA polymerase purified using aqueous two-phase extraction: Utilities of the enzyme in viral diagnosis

Abstract

HighlightsSso7d-Taq fusion protein purified using a single step of aqueous Two-Phase Extraction (ATPE) is >95% pure and is active.The S-Taq protein has higher thermostability and detergent tolerance over regular Taq polymerase and can be used for PCR's from direct whole blood.The PCR efficiency rate of S-Taq is higher than Taq polymerase and can be used to detect DNA viruses in a clinical setting efficiently.S-Taq can tolerate higher concentrations of magnesium ions and can be used for in-situ PCR’s.S-Taq can be used to carry out PCR’s of bacterial recombinants directly from the overnight culture since it is resistant to inhibition to Luria Bertani broth. This unique property of S-Taq will enable researchers to screen recombinants without the need to isolate the plasmid DNA of recombinants. This would be a huge cost savings for companies engaged in molecular biology research involving PCR’s.

A Single Amino Acid Change to Taq DNA Polymerase Enables Faster PCR, Reverse Transcription and Strand-Displacement

“…From this result, and our results of faster PCR, we assume that TaqD732N also has increased processivity. We suspect that Sso7d-Taq has not been tested for RT activity nor LAMP activity, although it has recently been extensively compared to wild-type Taq for PCR ( Sundarrajan et al, 2018 ). It would not surprise us if this or other DNA polymerases that have been modified for increased processivity, or already have high processivity, also have reverse transcriptase and/or strand-displacement activity.…”