A full-length Na'/glucose cotransporter cDNA (SGLTl) from rabbit intestine was subcloned into the pMAMneo mammalian expression vector and transfected by Ca'+ precipitation into Madin-Darby canine kidney (MDCK) cells. Stable MDCK transfectants isolated after clonal isolation and selection in G418 exhibited dexamethasone-inducible Na'/glucose cotransport activity under regulation of the MMTV promoter of the vector. Transfectants expressed the recombinant 75 kDa Na'lglucose cotransporter subunit as shown by Western blot, and SGLTl mRNA as shown by Northern blot, but these were undetectable in untransfected MDCK cells. Over 93% of total recombinant transport activity was targeted to the apical membrane. This indicates that the primary amino acid sequence of SGLTl contains the information necessary to target this transporter to the apical membrane.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.