DNA-binding properties of GST-NP6. (A) The purity
of 50 ng of GST-NP6 eluted from a FPLC Mono-S column was determined by
PhastGel SDS-PAGE (Pharmacia) and silver staining (lane 2). A Western
blot of a similar aliquot of the same Mono-S fraction, with the use of
anti-GST antibody, is shown in lane 1. Protein molecular mass markers
are in lane 3. (B) EMSA of GST-NP6 with 10 fmol of ARS1
as the probe. Buffer only (lane 1) and GST only (1000 ng, lane 2) were
used as controls. GST-NP6 was added at 5 ng (lane 3), 10 ng (lane 4),
and 30 ng (lanes 5–9). For a competition assay, 0.2 μg of
poly(dA-dG):poly(dC-dT) (Poly dN; lane 6), 100 ng of the 283-bp
ARS1 (lane 7), 30 ng of the 210-bp ARS1-B
(lane 8), and 10 ng of the 70-bp ARS1-A (lane 9) were
also added.