Development of pXeDNA3.1-LacZ, a Dual-System Plasmid for Membrane Protein Expression in Xenopus and Mammalian Cells

Cross-species expression of proteins in mammalian cells and Xenopus oocytes remains a powerful strategy for functional studies. This can be facilitated by plasmids with dual promoters, enabling use in both systems without separate subcloning. We developed pXeDNA3.1-LacZ, a plasmid that supports expression in mammalian cells and oocytes. The construct combines a cytomegalovirus promoter for mammalian transcription with Xenopus β-globin untranslated regions to enhance oocyte translation, and incorporates XcmI-mediated TA cloning and blue/white selection. To validate it, we used human TRPV1 (HsTRPV1). The vector drove robust expression in HEK293G5A cells and Xenopus oocytes.pXeDNA3.1-LacZ streamlines assays and facilitates studies of membrane proteins.