Part:BBa_K4188006

afraGFP fused to PTS1 tag

Characterisation via fluorescent measurement

The afraGFP (BBa_K4188034) and afraGFP_PTS1 (BBa_K4188006) were inserted into a level 1 shuttle vector, the pSB1KY-HIS3. The ScpTEF promotor and the ScCYC1t terminator were added to the coding sequence.

100 µl of a 1:5 diluted Saccharomyces cerevisiae suspension derived from cultures incubated for two hours were added to a well plate primed with concanavalin A. After 10 min of incubation, the S. cerevisiae suspension was replaced with Sc-all medium and the cells were imaged using a Nikon N-SIM microscope with an exposure time of 100 ms at 2 % laser intensity. AfraGFP fluorescence was excited at 488 nm and emission was detected at 510 ± 15 nm. Subsequent image editing in B allowed for better visibility of the cell background. Scale bar 5 µm.

Figure 1: Fluorescence microscopy of S. cerevisiae cells exhibiting strong afraGFP signal in (A) cytosol (BBa_K4188034) and (B) peroxisomes.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]