Production of active mouse DNA polymerase delta in bacteria.

Hindges R, Hübscher U
Gene (1995), Volume 158, Page 241
PubMed entry


The entire cDNA encoding the large subunit of mouse DNA polymerase ...
The entire cDNA encoding the large subunit of mouse DNA polymerase delta (mPol delta; EC has been cloned and expressed in various bacterial expression systems. A soluble protein could only be obtained when mPol delta was produced as a glutathione S-transferase (GST) fusion protein and the incubation temperature of the expression strain was reduced to 30 degrees C. After purification over a glutathione-Sepharose column, the fractions containing the recombinant (re-) fusion protein showed both DNA Pol and 3'-->5' Exo activities. In situ activity gel analysis indicated that the Pol activity resides in the re-protein. This activity, however, was not stimulated by proliferating cell nuclear antigen (PCNA). Our data are discussed in the view of the findings of Goulian et al. [J. Biol. Chem., 265 (1990) 16402-16411] that the second mPol delta subunit, the 48-kDa protein, might play an important role in DNA Pol delta-PCNA interaction.




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