The inability in deleting a reasonably large (>100 bp) DNA region with existing oligonucleotide-based deletion mutagenesis methods led to the application of a highly efficient RE-mediated inverse PCR method for the deletion of large areas in abnormally large P. falciparum genes. The method incorporates unique restriction enzyme sites at the 5'-ends of inverse tail-to-tail primers. In the absence of unique restriction sites, alternative methods including DiSec/TriSec [25], which allows the generation of speci- fied sticky-ends, may be used or a restriction-independent method like the overlapping primer method should suf- fice. The method has proven to be invaluable in decipher- ing the involvement of parasite-specific inserts in structure-activity relationships of PfAdoMetDC/ODC and PfPdxK (manuscript in preparation).