Fig. 2.

Plasmid-based C-terminal epitope tagging. (A) Production scheme for C-terminal HA tagging construct for the RPB2 gene. See text for details. (B) Southern blot analysis of an RPB2-HA strain of Tetrahymena. Genomic DNA from both the wild-type B2086 and RPB2-HA (2)-3-4 strains were digested with BamHI (Ba in panel A) and BglII (Bg panel A). The southern blot was prepared using the DNA from these strains and was probed with the DNA fragment shown in panel A. The bands corresponding to both the wild-type RPB2 and the RPB2-HA are marked by arrowheads. (C) Rpb2p-HA localization in a vegetative Tetrahymena cell. Rpb2p was localized using an anti-HA antibody (magenta). DNA was counter-stained with DAPI (blue). The macro- and micronuclei are marked with “a” and “i”, respectively.