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