EGFP in S. cerevisiae
We are trying to express and detect EGFP protein in S. cerevisiae.
Gene sequence has been checked, and Nothern Blot analysis indicates
that the gene is transcribed, nevertheless, we are not able to
detect any fluorescence (fluorescence microscope, FACS,
Fluorometer). We decided to construct a strong positif control (EGFP
cloned in a multi-copy plasmid, under control of a strong inducible
promoter, and transformed in a protease deficient yeast strain), but
as previously, no fluorescence is detected. Cells have been grown on
liquid and solid medium, at 30C and at room temperature.
Has anybody same problems to detect EGFP or has someone some
indications which could help us ?
Thank you for your help.
bozic at britbio.co.uk