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Plasmodium falciparum: generation of a cDNA library enriched in sporozoite-specific transcripts by directional tag
D A Fidock1, T V Nguyen, J M Ribeiro
1Department of Molecular Biology and Biochemistry, University of California, Irvine, California, 92697, USA. dfidock@aecom.yu.edu
Abstract:
We have adapted the "directional tag subtractive hybridization" technique as a means of investigating stage-specific gene expression in Plasmodium falciparum. This technique utilizes unidirectional cDNA libraries cloned into separate lambda vectors and involves hydroxyapatite chromatographic separation of target antisense cDNA and driver sense strand cRNA followed by PCR amplification of cDNA sequences specific to the target stage. This technique enabled efficient subtraction of asexual blood stage sequences from a P. falciparum sporozoite cDNA library and led to identification of novel sporozoite sequences. This technique can be applied to study gene expression in parasite stages that are difficult to obtain routinely.