T. Wohl et al., DETECTION OF NONFUNCTIONAL OVEREXPRESSION GENE-PRODUCTS USING 2-DIMENSIONAL GEL-ELECTROPHORESIS WITH A NARROWED PH RANGE, Electrophoresis, 16(5), 1995, pp. 876-878
A commonly used technique in the analysis of yeast protein function is
the overexpression of cloned genes. In the case that overexpression d
oes not lead to an altered phenotype a stable synthesis of the protein
has to be demonstrated. Here an example is shown where overexpression
of the yeast HYP2 genes, coding for a hypusine-containing protein, wa
s alternatively analyzed by sodium dodecyl sulfate polyacrylamide gel
electrophoresis (SDS-PAGE) alone or by two-dimensional (2-D) gel elect
rophoresis, using the narrowed pH range of 4.5-5.4. The results of the
SDS-PAGE suggested a stable overproduction of HYP gene product, where
as 2-D analysis revealed an accumulation of nonfunctional protein isof
orms.