Skip navigation

Development and validation of four Leishmania species constitutively expressing GFP protein. A model for drug discovery and disease pathogenesis studies

Development and validation of four Leishmania species constitutively expressing GFP protein. A model for drug discovery and disease pathogenesis studies

Parbhu Patel, Asha, Deacon, Andrew and Getti, Giulia (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:10.1017/S0031182013001777)

[img] PDF (Publisher's PDF)
10651 GETTI_Development_Validation_Leishmania_2013.pdf - Published Version
Restricted to Registered users only

Download (586kB)

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 / Department / Research Group: Faculty of Engineering & Science
Faculty of Engineering & Science > Department of Life & Sports Sciences
Related URLs:
Last Modified: 07 Apr 2017 20:30
Selected for GREAT 2016: GREAT a
Selected for GREAT 2017: None
Selected for GREAT 2018: None
URI: http://gala.gre.ac.uk/id/eprint/10651

Actions (login required)

View Item View Item

Downloads

Downloads per month over past year

View more statistics