USE OF RANDOM AMPLIFIED POLYMORPHIC DNA (RAPD) TECHNIQUE IN INHERITANCE STUDIES OF PLASMODIUM-FALCIPARUM

Effectiveness of random amplified polymorphic DNA (RAPD), a technique using 1 10-base primer to amplify random segments of genomic DNA, and some of its possible uses were tested in the A+T-rich genome of Plasmo dium falciparum. The best concentrations of MgCl2, 60% G+C primer and DNA were determined to be 4.0 mM, 0.4 mu M, and 90-180 ng/15 mu l reac tion, respectively. Use of 30% G+C primers did not allow amplification to occur. Application of RAFD to DNA of parent and progeny clones fro m a P. falciparum crest showed that polymorphisms identified in the pa rentals and tracked in the progeny were inherited in a Mendelian fashi on and that RAPD-identified polymorphisms could be used as genetic mar kers. Some of these polymorphic markers were located on more than 1 ch romosome, whereas others were specific for a single chromosome. Two of these markers, each located on chromosome 3 of 1 of the parental para sites, were missing from 2 of the 18 progeny, suggesting that deletion s, or crossover events had occurred. RAPD markers also identified a hi gher number of nonparental-type progeny than expected, thus confirming previous observations for high genetic variablity in malaria parasite s.