A cDNA clone which encodes a monocarboxylate transporter (ratMCT1) was isolated from a rat small intestinal cDNA library, which was screened by using full-length MCT1 cDNA of Chinese hamster ovary cells. The ratMCT1 cDNA was sequenced and predicted a protein of 494 amino acids with twelve potential transmembrane domains. The amino acid sequence showed 93.1% and 84.6% identity to the hamster and human monocarboxylate transporters, respectively. When expressed in Xenopus laevis oocytes, the ratMCT1 cRNA caused a significant increase in the uptake of radiolabeled lactic acid. Poly(A)+ RNA transcripts hybridizing to the ratMCT1 cDNA were detected in rat brain, heart, kidney, lung, muscle and brain capillaries. These results indicate that MCT1 contributes to pH-dependent and carrier-mediated transport of monocarboxylic acids in many tissues, not just in the small intestine.