GFP expressing stable cell lines

Muralikrishna Vemula vemula at helix.mgh.harvard.edu
Fri Mar 29 17:42:22 EST 2002


Hello,

re: problems with GFP expression

I cloned a fragement of DNA that contains 4 copies of NFkB responsive
element-TK promoter-d2EGFP fusion from plasmid pNFkB-d2EGFP (sold by
Clontech) into pd2GFP (again sold by Clontech) that has Neo marker in it.
I linearized the plasmid by using an enzyme that cuts approximately 100
bases ahead of the NFkB responsive elements and transfected HepG2
(hepatocyte) cells. I then established a stable cell line that is 
resistant to 500 ug/mL G418 for several generations now. On inducing these
cells with 100 ng/mL of IL1 for as long as 4-6 hours, I dont see any GFP
fluorescence. Does anybody have any thoughts why it is not working? Thanks
for any advice.

Murali Vemula

___________________________________________________________________
Murali Vemula, Ph.D
Research Fellow
Harvard Medical School/Shriners Hospital
51 Blossom Street
Boston, MA 02114
617-371-4927 (phone)
617-371-4951 (fax)

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