Supplementary Figure S4. Simultaneous multiple
protein expression from a single vector for Dictyostelium.
(A) Scheme of tandem GOI alignment using
seamless cloning (In-fusion cloning). After linearization of the vector
containing GOI1 with ApaI (whose recognition site is not present in 99.5% of
the Dictyostelium genes), a PCR product containing GOI2 with a promoter
(orange) and terminator (cyan, shaded) was inserted. (B) A γ-tubulin–GFP
expressing cell. G418 was used for selection. (C) A γ-tubulin–GFP and
mRuby3–histone H1 expressing cell. Only G418 was used for selection. The right
side of each image superimposed to the phase-contrast image. Bars, 10 µm.