Firefly luciferase-expressing U251 and PBT-017 (U251.ffluc, PBT-017.ffluc) and
DsRed.U251 were provided by Dr.
coli
dsRed. PBMO/CBMO were seeded in a density of 5 x 106 cells/[cm.sup.2] to the slide and were incubated for 23 h.
The enhanced green fluorescent protein (eGFP) transgene was replaced with the
DsRed transgene following Cre recombination.
Transgenic cells expressed
dsRed fluorescent protein and thus were identified via fluorescence microscopy 72 hr post-transduction.
Previously, via the somatic cell nuclear transfer (SCNT) technique, we successfully generated transgenic Guangxi Bama mini-pigs carrying a fluorescent protein (
DsRed) reporter gene regulated by the 2.2-kb hGFAP promoter (hGFAP-DsRed) (Zhu et al., 2016b, c).
Cells were observed on a Leica confocal microscope TCS SP8 using the following excitation and emission wavelengths: GFP (excitation 488 nm; emission 509 nm), mCherry (excitation 587 nm; emission 610 nm),
dsRed (excitation 560 nm; emission 592 nm), and MitoTracker Red (excitation 578 nm; emission 600 nm).
The T2A peptide is posttranslationally cleaved, releasing native ZF [gamma]1 and the fluorescent
DsRed reporter in equal molar concentrations (Prabhudesai et al.
ECFP and
DsRed were mostly shown in cytoplasmic (with a punctuate pattern) in dots (Fig.
By 2002, Robert Campbell, working in Roger Tsien's laboratory at the University of California, San Diego, had developed a monomeric form of
DsRed by subjecting it to an arduous process of structure-guided directed evolution (3).
There was more
dsRED fusion protein being coded and detected in the sh500 group (Figure 4(a2)), indicating that sh500 had weaker silencing effect on goat PPARG.
Optimal excitation is offered for red fluorescent proteins such as mCherry and
DsRed, which are being increasingly used in research labs.
Hep2G cells transfected with EGFP and
DsRed exhibited a bright-green and yellow fluorescence when viewed under a GFP and FITC filter, respectively.