Split mCherry as a new red bimolecular fluorescence complementation system for visualizing protein–protein interactions in living cells

A disulphide-linked heterodimer of TWIK-1 and TREK-1 mediates passive conductance in astrocytes

“…However, after careful examination of the report, we see that the discrepancy arises from technical limitations of the FRET and BiFC techniques. Although these techniques are powerful tools that permit visualization of protein-protein interactions 45,46 , the distance between two fluorescent proteins within the constructs is the most important factor for achieving a positive signal 24,47 . Indeed, our BiFC experiments showed that some combination of constructs (TWIK-1-VC þ VN-TREK-1 or TREK-1-VC þ VN-TWIK-1) gave a strong fluorescence signal (Fig.…”

“…To do this, we used the bimolecular fluorescence complementation (BiFC) technique 23 , which allows visualization of two independent proteins in close spatial proximity (a minimal distance of 7 nm (ref. 24) and TREK-1, whose N-and C terminus were fused with one complementary half of split Venus fluorescent protein, either the N-terminal half (VN) or the C-terminal half (VC), and then both were transfected into COS-7 cells (Fig. 2c).…”

Gateway Vectors for Plant Genetic Engineering: Overview of Plant Vectors, Application for Bimolecular Fluorescence Complementation (BiFC) and Multigene Construction

“…However, other fluorescent proteins, BFP (Hu & Kerppola, 2003), CFP (Kodama & Wada, 2009;Lee et al, 2008), GFP (Hu et al, 2002;Kodama & Wada, 2009), Venus, (Lee et al, 2008), Cerulean (Lee et al, 2008), DsRed-monomer (Kodama & Wada, 2009), mRFP1 (Jach et al, 2006), mCherry (Fan et al, 2008), and a far-red fluorescent protein, mLumin (Chu et al, 2009), have reportedly been useful for BiFC assay. We adopted CFP, GFP, YFP and mRFP1 to generate vectors ( Figure 9B), and verified their usefulness for detection of protein-protein interactions ( Figure 10B-E).…”

Improvement of a Venus-Based Bimolecular Fluorescence Complementation Assay to Visualize bFos-bJun Interaction in Living Cells

“…[2][3][4][5][6][7][8][9] Venus, a bright-yellow variant of GFP, efficiently matures at physiological temperature, and the intensity of fluorescence is higher than EYFP. 10) Thus Venus is suitable for the visualization of cellular structures.…”