A method to amplify and express malaria parasite genes containing an intron without cDNA synthesis.

Abstract Background The amplification of eukaryotic genes, which contain introns for expression purposes, is very tedious and expensive. The only available lab method is through cDNA synthesis, which has a very low success rate in generating a pure gene fragment lacking any introns. Methods The current paucity of any alternative lab method directed us to devise a strategy to amplify a P. falciparum RPL12mito gene (PfRPL12mito) containing an intron through three sequential PCR steps using gDNA and primers with a minimum 15 bp 5′-overhang for the full-length protein expression. Results Sanger sequencing confirmed the absence of an intronic segment and a continuous exonic region in the amplified gene fragment. The gene was successfully expressed as recombinant protein with a 6xHis tag. Conclusions This method provides a novel and cost-effective approach to amplify intron-containing eukaryotic genes, including those from the malaria parasite, directly from genomic DNA for expression purposes.