Ashutosh, * ; Gupta, Suman ; Ramesh, * ; Sundar, Shyam ; Goyal, Neena (2005) Use of Leishmania donovani field isolates expressing the luciferase reporter Gene in in vitro drug screening Antimicrobial Agents and Chemotherapy, 49 (9). pp. 3776-3783. ISSN 0066-4804
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Official URL: http://aac.asm.org/content/49/9/3776.short
Related URL: http://dx.doi.org/10.1128/AAC.49.9.3776-3783.2005
Abstract
Currently available primary screens for the selection of candidate antileishmanial compounds are not ideal. These techniques are time-consuming, laborious, and difficult to scale and require macrophages, which limit their use for high-throughput screening. We have developed Leishmania donovani field isolates that constitutively express the firefly luciferase reporter gene (luc) as a part of an episomal vector. An excellent correlation between parasite number and luciferase activity was observed. luc expression was stable, even in the absence of drug selection, for 4 weeks. The transfectants were infective to macrophages, and intracellular amastigotes exhibited luciferase activity. The suitability of these recombinant field isolates for in vitro screening of antileishmanial drugs was established. The luciferase-expressing sodium stibogluconate-resistant cell lines offer a model for the screening of compounds for resistance. The system is in routine use at the Central Drug Research Institute, Lucknow, India, for high-throughput screening of newly synthesized compounds.
Item Type: | Article |
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Source: | Copyright of this article belongs to American Society for Microbiology. |
ID Code: | 94556 |
Deposited On: | 16 Nov 2012 11:02 |
Last Modified: | 16 Nov 2012 11:02 |
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