MedicGo
Use of split-dihydrofolate reductase for the detection of protein-protein interactions and simultaneous selection of multiple plasmids in Plasmodium falciparum.
Metadata
Journalmolecular and biochemical parasitology1.571Date
2020 Jun 04
4 months ago
Type
Journal Article
Volume
2020-Jul / 238 : 111292
Author
Levray YS 1, Berhe AD 2, Osborne AR 3
Affiliation
  • 2. Pomona College, 333 N. College Way, Claremont, CA 91711, United States.
  • 3. Institute of Structural and Molecular Biology, University College London, Gower Street, London, WC1E 6BT, United Kingdom; Institute of Structural and Molecular Biology, Department of Biological Sciences, Birkbeck, Malet Street, London, WC1E 7HX, United Kingdom. Electronic address: [email protected]
Doi
PMIDMESH
Abstract
Defining protein-protein interactions is fundamental to the understanding of gene function. Protein-fragment complementation assays have been used for the analysis of protein-protein interactions in various organisms. The split-dihydrofolate reductase (DHFR) protein-fragment complementation assay utilises two complementary fragments of the enzyme fused to a pair of potentially interacting proteins. If these proteins interact, the DHFR fragments associate, fold into their native structure, reconstitute their function and confer resistance to antifolate drugs. We show that murine DHFR fragments fused to interacting proteins reconstitute a functional enzyme and confer resistance to the antifolate drug WR99210 in Plasmodium falciparum. These data demonstrate that the split-DHFR method can be used to detect in vivo protein-protein interactions in the parasite. Additionally, we show that split-DHFR fragments can be used as selection markers, permitting simultaneous selection of two plasmids in the presence of a single antifolate drug. Taken together, these experiments show that split-DHFR represents a valuable tool for the characterisation of Plasmodium protein function and genetic manipulation of the parasite.
Keywords: Co-Transfection Malaria Plasmodium transfection Protein-Fragment complementation assay Selection marker Split-Dihydrofolate reductase
Fav
Like
Download
Share
Export
Cite
1.6
Mol Biochem Parasitolmolecular and biochemical parasitology
Metadata
LocationNetherlands
FromELSEVIER

No Data

© 2017 - 2020 Medicgo
Powered by some medical students