In Article <9603012056.AA00236 at med.unc.edu>, marilee at MED.UNC.EDU (Marilee
Shelton) wrote:
>Does anyone have any experience using GFP with retrovirus vectors
>or with an expression system based on a single copy of the gene?
>I am concerned that the fluorescence may not be detectable from a single
>integrated copy of the GFP gene. Thanks for any assistance.
>>Marilee Shelton
>>marilee at med.unc.edu>CB#7365, FLOB
>University of North Carolina
>Chapel Hill, NC 27510-7365
>919-966-5035 (phone)
>919-966-5640 (fax)
We've seen expression in colonies after a few weeks of selection, so the DNA
should be integrated. However, we haven't tried to determine copy number,
so there could be multiple copies of GFP in these cells. If you are
concerned about sensitivity, try using humanized GFP (available from
Clonetech). We seem to get much better glowing with this version in 293
cells than with either wild type or with an S65T version that retains
jellyfish codons. I've also heard that there may be a humanized version
available from BRL.
Stu Kuhstoss
