An 8.0-kb EcoRI fragment of the human immunoglobulin (Ig) lambda light chain locus carrying the Ke-Oz- and Ke-Oz+ constant region genes and flanking sequences was studied for the presence of a transcriptional enhancer. Two types of assays were used. In the first, we measured the ability of recombinant plasmids carrying the 8.0-kb Ig fragment covalently linked to the aminoglycoside phosphotransferase (aph) gene to transform mouse myeloma or mouse fibroblast cells to geneticin resistance. In the second, we used RNA spot and Northern blot hybridization analyses to determine the relative levels of aph specific RNA transiently expressed after DNA transfection. In fibroblast cells, the transformation frequencies were independent of the presence of the Ig fragment in the vector and there was no difference in the level of transient expression of the aph gene. In myeloma cells, the Ig fragment enhanced at least 10-fold both the number of transformants and the level of aph gene expression over that obtained with vector alone. These results indicate the presence of a tissue-specific transcriptional enhancer in the 8.0-kb EcoRI fragment of the human Ig C lambda locus.