ANALYSIS OF PYRUVATE KINASE-DEFICIENCY MUTATIONS THAT PRODUCE NONSPHEROCYTIC HEMOLYTIC-ANEMIA

The intron sequences of the human L-type pyruvate kinase gene (PKLR) w ere determined by using primers selected from the known cDNA sequence. Oligonucleotide primers for these determined intron sequences were us ed to sequence the exons. When this technique was applied to the DNA o f 10 unrelated patients with pyruvate kinase deficiency, the following eight different mutations in the coding region were detected: del391- 393, A401, C464, G721, A1076, T1456, T1484, A1529. The A1529 mutation was found repeatedly in unrelated individuals, even in the homozygous state. The context with respect to a polymorphism at nt 1705 was compa tible with a single origin for this mutation, and it may represent a b alanced polymorphism. In normal subjects, five differences from the pu blished cDNA sequence were documented.