A water-soluble PEG derivative having both amino and carboxylic acid side chains (PEG-AC) was synthesized and explored for its transcription- and transfection-enhancing activity. PEG-AC could be deposited onto the surface of DNA/polycation complexes to form a ternary complex with slightly negative surface potential. PEG-AC-coating on the plasmid/PEI complexes obviously enhanced their transcriptional activity, and 31-fold higher consumption of UTP was observed. Amphoteric PEG-AC would loosen the tightly compacted DNA/PEI complex and facilitate the approach of transcriptional factors. PEG-AC also evidently improved the transgene expression level on the cultured CHO cells.