We have cloned the cDNA of the rat liver 13k protein, that has glutathione binding activity, from a rat liver cDNA library. The nucleotide sequence of this cDNA predicts a protein of 115 amino acids. This protein has 99.1%, 89.6% and 73.0% homology with mouse, human and chicken macrophage migration inhibitory factor (MIF), respectively. This indicates that the rat liver 13k protein is homologue of the MIF. The N-terminal 25 amino acids show 35% sequence similarity with that of the rat glutathione transferase Yb subunit. Although the remaining C-terminal sequence has no sequence identity with glutathione transferase Yb subunit, about 50% homology was found, if conservative substitutions and gaps were taken into account. Northern blot analysis indicates that this gene is expressed in a wide variety of organs including brain, spleen, liver, muscle and kidney. Southern blot analysis suggests that the MIF gene has many pseudo-genes or/and closely related genes.