Figure 8.
In situ detection of trans-activator of transcription (TAT)-driven phage-mediated green fluorescent protein (GFP) expression in cultured cells. COS-1 cells (2.5×104) were incubated for 6 hours at 37°C, with 500 μL of medium containing 2.5×1010 pfu of TAT GFP phages (A,B), 2.5×109 pfu of TAT GFP phages (C,D), or 2.5×1010 pfu of wild-type GFP phages (E,F). pfu=plaque-forming units. After 48 h, the cells were counterstained with 4′,6-diamidine-2-phenylindole HCl and examined by fluorescence microscopy, with a GFPA cube (A,C,E) or a WU cube (B,D,F), as described in the report. G) Preparation of phage λ particles displaying foreign peptides. DNA is packaged through recognition of the COS site into the phage λ prohead, which consists of E protein. Next, the chimeric D protein with the foreign peptide is assembled onto the prohead, followed by binding of the head to the tail.[37]