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Additional file 8. Chimeric rodent parasite lines pb-pvcsp(vk210) and pb-pvcsp(vk247) are able to produce salivary gland sporozoites that are infectious to mice. A. Upper panel, left: Light microscope pictures of wild type P. berghei (WT) and pb-pvcsp oocyst at days 7, 10 and 14 after feeding of An. stephensi mosquitoes. Black arrows indicate oocyst with sporozoite formation and white arrows indicate degenerated, vacuolated oocyst without sporozoite formation (scale bar, 10 µm). Upper panel, right, Percentage of degenerated oocyst in An. stephensi mosquitoes (n = 5) at day 14 after feeding (***P =

Additional file 8. Chimeric rodent parasite lines pb-pvcsp(vk210) and pb-pvcsp(vk247) are able to produce salivary gland sporozoites that are infectious to mice. A. Upper panel, left: Light microscope pictures of wild type P. berghei (WT) and pb-pvcsp oocyst at days 7, 10 and 14 after feeding of An. stephensi mosquitoes. Black arrows indicate oocyst with sporozoite formation and white arrows indicate degenerated, vacuolated oocyst without sporozoite formation (scale bar, 10 µm). Upper panel, right, Percentage of degenerated oocyst in An. stephensi mosquitoes (n = 5) at day 14 after feeding (***P =

Additional file 4. Growth of asexual blood stages and mCherry expression in blood stages. Growth of asexual blood stages of two chimeric P. falciparum parasite lines (pf-pvcsp(vk210) and pf-pvcsp(vk247), a P. falciparum line lacking expression of CSP (PfΔcsp) and P. falciparum wild type (WT) parasites. Parasites of the different cloned lines were cultured in semi-automated culture system for a period of 7 days. Cultures were initiated with a parasitaemia of 0.5%. Arrows indicate the dilution of the cultures with fresh red blood cells to have a final parasitaemia of 1%. B. mCherry-expressing blood stages of PfΔcsp parasites where the csp gene has been disrupted by insertion of an mCherry expression cassette (see Additional file 5 for details of the generation of PfΔcsp). Scale bar, 7 µm.

Additional file 3. Sequence analysis of the P. vivax csp genes introduced into P. falciparum. Long-range PCR fragments (see Fig. 1) were cloned into pET TOPO TA (Invitrogen) and sequenced. See Additional file 2 for primer sequences used for sequencing the complete fragment. A. LR-PCR fragment of parasites of pf-pvcsp(vk210). B. LR-PCR fragment of parasites of pf-pvcsp(vk247).

Additional file 1. Vector maps of the P. vivax csp genes introduced into P. falciparum. A. Vector maps of the different plasmids used to generate two chimeric P. falciparum parasite lines (pf-pvcsp) expressing P. vivax CSP(VK210) of CSP(VK247) and a P. falciparum line lacking expression of CSP (PfÎ csp). See â Methodsâ section for description and details of the generation of these plasmids.

Additional file 5. Degenerated oocysts of two chimeric P. falciparum parasite lines (pf-pvcsp) and a PfCSP knockout line (PfΔcsp). Light microscope pictures of degenerated oocysts at day 10 after feeding gametocytes to Anopheles stephensi mosquitoes. These oocysts are classified as degenerate based on the absence of sporozoite formation and vacuolated cytoplasm. See Fig. 2 for pf-pvcsp and wild type P. falciparum (WT) oocyst in which sporozoite formation occurred. No sporozoite formation was observed in PfΔcsp oocysts. Scale bar, 20 µm.

Additional file 4. Growth of asexual blood stages and mCherry expression in blood stages. Growth of asexual blood stages of two chimeric P. falciparum parasite lines (pf-pvcsp(vk210) and pf-pvcsp(vk247), a P. falciparum line lacking expression of CSP (PfΔcsp) and P. falciparum wild type (WT) parasites. Parasites of the different cloned lines were cultured in semi-automated culture system for a period of 7 days. Cultures were initiated with a parasitaemia of 0.5%. Arrows indicate the dilution of the cultures with fresh red blood cells to have a final parasitaemia of 1%. B. mCherry-expressing blood stages of PfΔcsp parasites where the csp gene has been disrupted by insertion of an mCherry expression cassette (see Additional file 5 for details of the generation of PfΔcsp). Scale bar, 7 µm.

Additional file 5. Degenerated oocysts of two chimeric P. falciparum parasite lines (pf-pvcsp) and a PfCSP knockout line (PfΔcsp). Light microscope pictures of degenerated oocysts at day 10 after feeding gametocytes to Anopheles stephensi mosquitoes. These oocysts are classified as degenerate based on the absence of sporozoite formation and vacuolated cytoplasm. See Fig. 2 for pf-pvcsp and wild type P. falciparum (WT) oocyst in which sporozoite formation occurred. No sporozoite formation was observed in PfΔcsp oocysts. Scale bar, 20 µm.

Additional file 1. Vector maps of the P. vivax csp genes introduced into P. falciparum. A. Vector maps of the different plasmids used to generate two chimeric P. falciparum parasite lines (pf-pvcsp) expressing P. vivax CSP(VK210) of CSP(VK247) and a P. falciparum line lacking expression of CSP (PfÎ csp). See â Methodsâ section for description and details of the generation of these plasmids.

Additional file 3. Sequence analysis of the P. vivax csp genes introduced into P. falciparum. Long-range PCR fragments (see Fig. 1) were cloned into pET TOPO TA (Invitrogen) and sequenced. See Additional file 2 for primer sequences used for sequencing the complete fragment. A. LR-PCR fragment of parasites of pf-pvcsp(vk210). B. LR-PCR fragment of parasites of pf-pvcsp(vk247).