Parbhu Patel, Asha, Deacon, Andrew and Getti, Giulia ORCID: 0000-0003-1402-8496 (2013) Development and validation of four Leishmania species constitutively expressing GFP protein. A model for drug discovery and disease pathogenesis studies. Parasitology, 141 (4). pp. 501-510. ISSN 0031-1820 (Print), 1469-8161 (Online) (doi:https://doi.org/10.1017/S0031182013001777)

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Abstract

Green fluorescent protein (GFP)-parasite transfectants have been widely used as a tool for studying disease pathogenesis in several protozoan models and their application in drug screening assays have increased rapidly. In the past decade, the expression of GFP has been established in several Leishmania species, mostly for in vitro studies. The current work reports generation of four transgenic parasites constitutively expressing GFP (L. mexicana, L. aethiopica, L. tropica and L. major) and their validation as a representative model of infection. This is the first report where stable expression of GFP has been achieved in L. aethiopica and L. tropica. Integration of GFP was accomplished through homologous recombination of the expression construct, pRib1.2αNEOαGFP downstream of the 18S rRNA promoter in all species. A homogenous and high level expression of GFP was detected in both the promastigote and the intracellular amastigote stages. All transgenic species showed the same growth pattern, ability to infect mammalian host cells and sensitivity to reference drugs than their wild type counterparts. All four transgenic Leishmania are confirmed as models for in vitro and possibly in vivo infections and represent an ideal tool for medium 31 throughput testing of compound libraries.

Item Type:	Article
Uncontrolled Keywords:	Leishmania aethiopica; Leishmania major; Leishmania Mexicana; 34 Leishmania tropica; flow cytometry, intracellular amastigote
Subjects:	R Medicine > RS Pharmacy and materia medica
Faculty / School / Research Centre / Research Group:	Faculty of Engineering & Science Faculty of Education, Health & Human Sciences > School of Human Sciences (HUM)
Related URLs:	Publisher Publisher Organisation
Last Modified:	09 Oct 2021 04:46
URI:	http://gala.gre.ac.uk/id/eprint/10651

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