Plasmodium falciparum: Generation of a cDNA library enriched in sporozoite-specific transcripts by directional tag subtractive hybridization

We have adapted the "directional tag subtractive hybridization" technique a s a means of investigating stage-specific gene expression in Plasmodium fal ciparum. This technique utilizes unidirectional cDNA libraries cloned into separate lambda vectors and involves hydroxyapatite chromatographic separat ion of target antisense cDNA and driver sense strand cRNA followed by PCR a mplification of cDNA sequences specific to the target stage. This technique enabled efficient subtraction of asexual blood stage sequences from a P. f alciparum sporozoite cDNA library and led to identification of novel sporoz oite sequences. This technique can be applied to study gene expression in p arasite stages that are difficult to obtain routinely. (C) 2000 Academic Pr ess.