Fig. 5

Verification of differentially spliced introns between Plasmodium y. nigeriensis NSM and Plasmodium y. yoelii 17XNL. DNA and mRNA samples were prepared from NSM and 17XNL parasites; 17XNL was used because YM parasite was not available in the laboratory in China. a, b RNA-seq read alignments showing a predicted intron and primer positions (arrows) from the PYYM_0205900 gene that has the same spliced intron in both NSM and YM (a) and amplification products using primers 5′-TGTCCATCAAATAATAAAGCTAAAATATATTCCTCTCA-3′ and 5′-TATAGTTAGATGTGTTTAATATTTAAGG-3′. Parasite names with ‘g’ indicate amplification products from genomic DNA, and those with ‘c’ were from cDNA. The results showed no DNA band in the cDNA preparations, suggesting lack of DNA contamination in the cDNAs. c, d RNA-seq alignments (c) and amplification products (d) from gene PYYM_0406600 using primers 5′-GTAAGAAATATACAACAATACTATTCCTTGGCAA-3′and 5′-CTCTCCCATTTTTAGGTATAAAAAATAACTAAATATG-3′, showing a much stronger sliced product (arrowhead) in 17XNL than in NSM. e, f RNA-seq alignments (e) and amplification products from gene PYYM_0710900 (f) showing differential spliced bands (arrowheads) between NSM and 17XNL parasites (primers: 5′-GATTTCTATTAGCTTTGTGAAGTC-3′ and 5′- TGTAATATATTATCGAAAGACGTG-3′). In addition to differential splicing, size polymorphism in genomic DNA between NSM and 17XNL was also detected