Highly efficient subcloning of rodent malaria parasites by injection of single merosomes or detached cells

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

[img] Text
Nat Protoc 2009 Stanway.pdf - Published Version
Restricted to registered users only
Available under License Publisher holds Copyright.

Download (314kB) | Request a copy

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.

Item Type:

Journal Article (Original Article)

Division/Institute:

08 Faculty of Science > Department of Biology > Institute of Cell Biology
08 Faculty of Science > Department of Biology > Institute of Cell Biology > Malaria

UniBE Contributor:

Limenitakis Stanway, Rebecca and Heussler, Volker

Subjects:

500 Science > 570 Life sciences; biology
500 Science

ISSN:

1754-2189

Publisher:

Nature Publishing Group

Language:

English

Submitter:

Volker Heussler

Date Deposited:

01 Jun 2016 14:20

Last Modified:

01 Jun 2016 14:20

Publisher DOI:

10.1038/nprot.2009.172

PubMed ID:

19745825

BORIS DOI:

10.7892/boris.81869

URI:

https://boris.unibe.ch/id/eprint/81869

Actions (login required)

Edit item Edit item
Provide Feedback