Fluorescent Plasmodium berghei sporozoites and pre-erythrocytic stages: a new tool to study mosquito and mammalian host interactions with malaria parasites


Natarajan, R; Thathy, V; Mota, MM; Hafalla, JC; Menard, R; Vernick, KD; (2001) Fluorescent Plasmodium berghei sporozoites and pre-erythrocytic stages: a new tool to study mosquito and mammalian host interactions with malaria parasites. Cellular microbiology, 3 (6). pp. 371-9. ISSN 1462-5814 DOI: https://doi.org/10.1046/j.1462-5822.2001.00117.x

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Abstract

To track malaria parasites for biological studies within the mosquito and mammalian hosts, we constructed a stably transformed clonal line of Plasmodium berghei, PbFluspo, in which sporogonic and pre-erythrocytic liver-stage parasites are autonomously fluorescent. A cassette containing the structural gene for the FACS-adapted green fluorescent protein mutant 2 (GFPmut2), expressed from the 5' and 3' flanking sequences of the circumsporozoite (CS) protein gene, was integrated and expressed at the endogenous CS locus. Recombinant parasites, which bear a wild-type copy of CS, generated highly fluorescent oocysts and sporozoites that invaded mosquito salivary glands and were transmitted normally to rodent hosts. The parasites infected cultured hepatocytes in vitro, where they developed into fluorescent pre-erythrocytic forms. Mammalian cells infected by these parasites can be separated from non-infected cells by fluorescence activated cell sorter (FACS) analysis. These fluorescent insect and mammalian stages of P. berghei should be useful for phenotypic studies in their respective hosts, as well as for identification of new genes expressed in these parasite stages.

Item Type: Article
Keywords: Animals, Anopheles, Antigens, Differentiation, Base Sequence, Green Fluorescent Proteins, Luminescent Proteins, Molecular Sequence Data, Plasmodium berghei, Recombinant Fusion Proteins, Animals, Anopheles, parasitology, Antigens, Differentiation, Base Sequence, Green Fluorescent Proteins, Luminescent Proteins, Molecular Sequence Data, Plasmodium berghei, cytology, genetics, growth & development, Recombinant Fusion Proteins
Faculty and Department: Faculty of Infectious and Tropical Diseases > Dept of Immunology and Infection
Research Centre: Malaria Centre
PubMed ID: 11422080
Web of Science ID: 169156100002
URI: http://researchonline.lshtm.ac.uk/id/eprint/9819

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