Dicistronic vectors utilizing the internal ribosomal entry site sequence of poliovirus as the intercistronic region were constructed for gene expression in mammalian cells. We have developed two monocistronic expression vectors which facilitate the creation of dicistronic expression plasmids. The dicistronic expression plasmids encode transcription units which allow the coordinated translation of the two genes. Using internal luciferase and secreted alkaline phosphatase, we show the correlated expression of both reporter genes and expression levels comparable to those achieved by the respective monocistronic expression vectors.