Figure 3From: A fluorescent cassette-based strategy for engineering multiple domain fusion proteinsFluorescence screening, protein purification and subcellular localization. (A) Fluorescence screening in bacterial colonies using the leak expression of the target protein fused with Venus. A non-fluorescent colony is identified by arrow 1; a fluorescent colony, by arrow 2. (B) The purification of the 6xHis-Venus-GST construct (identified by the arrow). Lane MW is the molecular weight marker; lane 1, the cell lysate; lane 2, the elusion from the Ni-NTA column; lane 3, the elusion from the GST column. All vectors were transfected into COS-7 cells for fluorescence imaging. The 10× magnification of (C) the cytoplasmic distribution of the 6xHis-Venus-GST and (E) the endoplasmic distribution of Venus N-terminally fused with the interleukin-4 leader sequence and C-terminally fused with the KDEL retention signal. (D) The 40× magnification of the nucleolar distribution of Venus N-terminally fused with the HIV Tat domain.Back to article page