Identification and recombinant expression of glyceraldehyde-3-phosphate dehydrogenase of Plasmodium falciparum

The gene coding for the cytosolic glyceraldehyde-3-phosphate dehydrogenase (GAPDH; EC 1.2.1.12) was isolated from Plasmodium falciparum. The gene cont ains 1 intron and the AST content is characteristic for the codon usage of P. falciparum. The predicted open reading frame codes for 337 amino acids ( 36 651 Dal and is 63.5% identical to the human erythrocytic GAPDH. GAPDH se quences from several field isolates of P. falciparum displayed 100% conserv ation. Phylogenetic analysis supports the hypothesis that dinoflagellates a nd Plasmodium are closely related. The protein encoded by the pfGAPDH was e xpressed recombinantly in Escherichia coli and exhibited enzymatic activity with NAD(+) but not with NADP(+) as cofactor. Antiserum raised against the recombinantly expressed enzyme detected specifically all developmental sta ges of cultured P. falciparum blood-stage parasites, (C) 2000 Elsevier Scie nce B.V. All rights reserved.