Figure 3. misGvA by yeast extracts producing variants of Pol ι.

A. misGvA activity of cell extracts of yeasts producing GST-Pol ι variants was estimated by “misGvA” in 8-min reactions in the presence of 0.15 mM Mn²⁺ and 0.25 mM Mg²⁺ divalent metal ions at template 1 as described in “Materials and Methods.” Two combinations of equal concentrations of 1 mM dNTPs have been used: 1) all dNTPs and 2) dGTP and dATP. B. Same with template 2. B. Western-blot analysis of the production of GST-tagged Pol ι catalytically compromised variants in yeast cells extracts. Extracts containing 40 µg of total protein were separated on 8% polyacrylamide-SDS gel, transferred to a PVDF membrane and probed with polyclonal anti-GST antibodies. Cross-reacting proteins were visualized according to the Western Breeze Chromogenic Anti-Rabbit Kit procedure (Invitrogen).