The FLT3 gene encodes a tyrosine kinase receptor that regulates proliferation and differentiation of hematopoietic cells. Recently, the internal duplication of FLT3 has been observed in hematological malignancies, suggesting the involvement of these mutations in leukemogenesis. The authors analyzed the expression of FLT3 mRNA and the incidence of its internal tandem duplication in normal hematopoietic and blood samples by reverse transcription-polymerase chain reaction (RT-PCR). Mononuclear cells (MNCs) in cord blood and bone marrow highly expressed FLT3 mRNA, whereas MNCs and polymorphonuclear cells (PMNs) in peripheral blood showed low or undetectable levels of FLT3. When the ratio of FLT3/beta-actin PCR products was calculated, the level of FLT3 mRNA expression was significantly higher in cord blood MNCs (n = 42) than that in peripheral MNCs (n = 14) or PMNs (n = 10). Although several PCR bands with different sizes were observed, no internal tandem duplication of FLT3 was detected in these normal blood samples. These findings indicate that the expression of FLT3 is lineage specific and consistently decreases during hematopoietic differentiation. The internal duplication of FLT3 is restricted in hematological malignancies and may occur at a specific stage in leukemogenesis.