A Bifunctional Fusion Enzyme with DNA polymerase and dUTPase Activities

Construction of a fusion enzyme by recombinant DNA technology could be useful in obtaining more efficient bifunctional enzymes. However, two open reading frames in tandem often interfere with each other's functions. We have previously demonstrated that when a relatively smaller dUTPase enzyme is tagged on either side of a thermostable DNA polymerase it did not affect polymerization function. Here, we will further reveal that both the enzymatic domains of these fusion enzymes also do not muddle each other's functions indeed. Furthermore, both of these affinity purified fusion proteins appeared to be ‘undisruptable dimers’ in denaturing protein gels and as confirmed by MALDI‐MS/MS analyses. Support or Funding Information Faculty Research Support Fund from the Office of Sponsored Programs at the University of Houston‐Clear Lake, Texas.