Connexin-43 (Cx43) containing giant liposomes (GL) were prepared by a baculovirus expression liposome fusion method Recombinant budded viruses expressing Cx43 were prepared and then fused with GLs containing DOPG/DOPC at pH 4 5 Connexon formation on the GL membrane was observed by transmission electron microscope Hydrophilic fluorescent dye transfers were observed through a Cx43-mediated pathway not only between Sf9 (Spodoptera frugiperda) cells with Cx43 but also from giant Cx43 liposomes to Cx43-expressing U2OS cells (human osteosarcoma cell) The functional connexin-containing liposome is expected to be useful for cellular cytosolic delivery systems The original orientation and function of Cx43 was maintained after integration into the liposomes The liposome fusion method will create new opportunities as a tool for analysis of channel membrane proteins Biotechnol Bioeng 2010,107 836-843 (C) 2010 Wiley Periodicails, Inc