Figure 1. Expression and purification of Pol ν-77.


(A) Schematic representation of the H. sapiens full-length and Pol ν-77 proteins. Diagram depicts an N-terminal region (NTR), of unknown function (white) and a C-terminal portion that encodes DNA polymerase activity. Conserved motifs in the polymerase domain are numbered 1, 2a, 2b and 3–6 (white boxes). Aspartate residues located in motif 3 that are important for DNA polymerase activity are represented as black squares. The extreme C-terminal region (amino acids 864 to 900, black) contains a proline rich tail. (B) Pol ν-77 samples were prepared as described in Materials and Methods: C, crude extract; P, pellet with insoluble fraction; S, soluble fraction; E, eluate of purified soluble Pol ν-77 fraction after Ni-NTA agarose (Qiagen) chromatography; M, molecular weight markers.