IRES sequence - pIRES2-EGFP doen't work (Jul/31/2003 )

Hello,

I subclone my gene (steroid receptor) from pEGFPC1 (Bgl2/Hind3) before the ires sequence in pIRES2-EGFP (Bgl2/Xma1).

I transfect this construction, in COS-7 DMEM + charcoal stripped FBS, with betagal control and reporter vector.

Betagal control is OK, my receptor is OK but i don't see fluo ?

Can you help me ?

Thanks a lot.

NB : the construction pEGFPC1-receptor is OK (fonction+fluo)

have you checked for the presence of a polyadenylation signal after the receptor you cloned in? If present this would stop the production of a bicistronic message by the IRES.