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Stable expression of Escherichia coli {beta}-glucuronidase A (GusA) in Giardia lamblia: application to high-throughput drug susceptibility testing


Müller, J; Nillius, D; Hehl, A B; Hemphill, A; Müller, N (2009). Stable expression of Escherichia coli {beta}-glucuronidase A (GusA) in Giardia lamblia: application to high-throughput drug susceptibility testing. Journal of Antimicrobial Chemotherapy, 64(6):1187-1191.

Abstract

Objectives In order to create a suitable model for high-throughput drug screening, a Giardia lamblia WB C6 strain expressing Escherichia coli glucuronidase A (GusA) was created and tested with respect to susceptibility to the anti-giardial drugs nitazoxanide and metronidazole. Methods GusA, a well-established reporter gene in other systems, was cloned into the vector pPacVInteg allowing stable expression in G. lamblia under control of the promoter from the glutamate dehydrogenase (gdh) gene. The resulting transgenic strain was compared with the wild-type strain in a vitality assay, characterized with respect to susceptibility to nitazoxanide, metronidazole and-as assessed in a 96-well plate format-to a panel of 15 other compounds to be tested for anti-giardial activity. Results GusA was stably expressed in G. lamblia. Using a simple glucuronidase assay protocol, drug efficacy tests yielded results similar to those from cell counting. Conclusions G. lamblia WB C6 GusA is a suitable tool for high-throughput anti-giardial drug screening.

Objectives In order to create a suitable model for high-throughput drug screening, a Giardia lamblia WB C6 strain expressing Escherichia coli glucuronidase A (GusA) was created and tested with respect to susceptibility to the anti-giardial drugs nitazoxanide and metronidazole. Methods GusA, a well-established reporter gene in other systems, was cloned into the vector pPacVInteg allowing stable expression in G. lamblia under control of the promoter from the glutamate dehydrogenase (gdh) gene. The resulting transgenic strain was compared with the wild-type strain in a vitality assay, characterized with respect to susceptibility to nitazoxanide, metronidazole and-as assessed in a 96-well plate format-to a panel of 15 other compounds to be tested for anti-giardial activity. Results GusA was stably expressed in G. lamblia. Using a simple glucuronidase assay protocol, drug efficacy tests yielded results similar to those from cell counting. Conclusions G. lamblia WB C6 GusA is a suitable tool for high-throughput anti-giardial drug screening.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:05 Vetsuisse Faculty > Institute of Parasitology
04 Faculty of Medicine > Institute of Parasitology
Dewey Decimal Classification:570 Life sciences; biology
610 Medicine & health
600 Technology
Language:English
Date:2009
Deposited On:17 Nov 2009 08:22
Last Modified:05 Apr 2016 13:33
Publisher:Oxford University Press
ISSN:0305-7453
Publisher DOI:10.1093/jac/dkp363
PubMed ID:19820251
Permanent URL: http://doi.org/10.5167/uzh-24107

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