Highly efficient subcloning of rodent malaria parasites by injection of single merosomes or detached cells
Data(s) |
2009
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Resumo |
This protocol describes a method for obtaining rodent Plasmodium parasite clones with high efficiency, which takes advantage of the normal course of Plasmodium in vitro exoerythrocytic development. At the completion of development, detached cells/merosomes form, which contain hundreds to thousands of merozoites. As all parasites within a single detached cell/merosome derive from the same sporozoite, we predicted them to be genetically identical. To prove this, hepatoma cells were infected simultaneously with a mixture of Plasmodium berghei sporozoites expressing either GFP or mCherry. Subsequently, individual detached cells/merosomes from this mixed population were selected and injected into mice, resulting in clonal blood stage parasite infections. Importantly, as a large majority of mice become successfully infected using this protocol, significantly less mice are necessary than for the widely used technique of limiting dilution cloning. To produce a clonal P. berghei blood stage infection from a non-clonal infection using this procedure requires between 4 and 5 weeks. |
Formato |
application/pdf |
Identificador |
http://boris.unibe.ch/81869/1/Nat%20Protoc%202009%20Stanway.pdf Limenitakis Stanway, Rebecca; Graewe, Stefanie; Rennenberg, Annika; Helm, Susanne; Heussler, Volker T. (2009). Highly efficient subcloning of rodent malaria parasites by injection of single merosomes or detached cells. Nature protocols, 4(10), pp. 1433-1439. Nature Publishing Group 10.1038/nprot.2009.172 <http://dx.doi.org/10.1038/nprot.2009.172> doi:10.7892/boris.81869 info:doi:10.1038/nprot.2009.172 info:pmid:19745825 urn:issn:1754-2189 |
Idioma(s) |
eng |
Publicador |
Nature Publishing Group |
Relação |
http://boris.unibe.ch/81869/ |
Direitos |
info:eu-repo/semantics/restrictedAccess |
Fonte |
Limenitakis Stanway, Rebecca; Graewe, Stefanie; Rennenberg, Annika; Helm, Susanne; Heussler, Volker T. (2009). Highly efficient subcloning of rodent malaria parasites by injection of single merosomes or detached cells. Nature protocols, 4(10), pp. 1433-1439. Nature Publishing Group 10.1038/nprot.2009.172 <http://dx.doi.org/10.1038/nprot.2009.172> |
Palavras-Chave | #570 Life sciences; biology #500 Science |
Tipo |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion PeerReviewed |