Plasmodium falciparum merozoite surface protein (MSP)-2, is a polymorphic protein whose variable regions define two allelic families, the 3D7/IC-1 and FC27/D10 families. The gene encoding MSP-2 is located on chromosome 2 immediately 3' of the gene encoding merozoite surface protein-5 (MSP-5) with a 1096 bp intergenic region that presumably contains the MSP-2 promoter. Here we present characterization of the MSP-2 promoter using transient and stable transfection of P. falciparum. The mRNA transcription initiation site was mapped to a position 256 bp upstream of the MSP-2 translation start site. The ability of the intergenic region between MSP-5 and MSP-2 to promote the expression of chloramphenicol acetyl transferase (CAT) has been tested using a series of nested deletions in transient transfection experiments. The minimal region required for CAT expression has been defined and putative regulatory elements delineated. These nested deletions were used for heterologous expression of an FC27 family MSP-2 allele in the 3D7 allelic background in transfected 3D7 lines. In each case, the transgenic P. falciparum lines generated co-express both 3D7 and FC27 allelic forms of MSP-2 at the merozoite surface. These results have identified the functional promoter for MSP-2.