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.
Tomo Kondo was supported by the Japan Society for the Promotion of Science Research Fellowships for Young Scientists. This research was supported by the Japan Society for the Promotion of Science [KAKENHI Grant Number 16J08310 and 19K15809 to TK]. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. No writing assistance was utilized in the production of this manuscript.