Efficiency of IRES (Internal Ribosome Entry Site) is high?

Jun-Ichi Aikawa jaikawa at UCSD.EDU
Mon Apr 20 18:12:34 EST 1998


To use EGFP (enhanced green fluorescent protein) as a reporter gene for
transfection, I put a fragment of synthetic intron and IRES (Internal
Ribosome Entry Site) prepared from pIRESneo (Clontech) upstream of EGFP.
When I expressed this plasmid transiently in CHO-K1 cells and analyzed its
fluorescence by FACS, expreesion of EGFP was detected, but much lower than
that of plasmid which has EGFP gene, but not IVS-IRES (pEGFP-N1). Is that
common in the case of use of IRES sequence? Otherwise, my strategy of
construction is wrong?  I welcome any comments and suggestions.

Thanks.

Jun-ichi Aikawa, Ph.D.
University of California, San Diego





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