Dear Bionetters,
During the last months it turned out that expression of GFP in
mamalien cells is a quite tricky story. Many scientist but definitley
not all did not get any fluorescence in cells like COS-1, COS-7, CHO,
NIH3T3, B-cells, HEPG2 and many others regardless they are trying to
express GFP or fusion proteins with it (any hints ?).
Some weeks ago I followed the discussion about the codon usage of
GFP-gene as it is used in the pGFP vectors. The general question was
wether a more "humanized" codon usage might solve the problem or not.
It propably might be helpfull in some cases.
Another maybe more important point might be the promotor especially in
the case of the pGFP-1 vector, which is thought to be a promotor test
vector. The general question (point of discussion) I want to post is
wether the distances between the CCAAT box and the TATA box to the
starting point of transcription are of major importance or not for the
expression of GFP in mamalien cells.
Is there anybody who does have some experiences or thoughts in this
point ?
Thanks to all answers,
Ulli Thoenes
e-mail: ulrich.thoenes at rhein-neckar.de
