Genomic Deoxyxylulose Phosphate Reductoisomerase (DXR) Mutations Conferring Resistance to the Antimalarial Drug Fosmidomycin in E. coli
Autor: | Ryan T. Gill, Eun Joong Oh, Gur Pines, Reilly G. Fankhauser, Carrie Eckert, Alaksh Choudhury, Marcelo C. Bassalo, Andrew D. Garst |
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Rok vydání: | 2018 |
Předmět: |
0301 basic medicine
Plasmodium falciparum 030106 microbiology Drug Resistance Biomedical Engineering Context (language use) Biology medicine.disease_cause Biochemistry Genetics and Molecular Biology (miscellaneous) Article Genome engineering Gene product Antimalarials 03 medical and health sciences Fosfomycin parasitic diseases Escherichia coli medicine CRISPR Clustered Regularly Interspaced Short Palindromic Repeats Gene Aldose-Ketose Isomerases Genetics General Medicine biology.organism_classification Fosmidomycin 030104 developmental biology Mutation Genetic Engineering Plasmids medicine.drug |
Zdroj: | ACS Synthetic Biology. 7:2824-2832 |
ISSN: | 2161-5063 |
Popis: | Sequence to activity mapping technologies are rapidly developing, enabling the generation and isolation of mutations conferring novel phenotypes. Here we used the CRISPR EnAbled Trackable genome Engineering (CREATE) technology to investigate the inhibition of the essential ispC gene in its native genomic context in Escherichia coli. We created a full saturation library of 33 sites proximal to the ligand binding pocket and challenged this library with the antimalarial drug fosmidomycin, which targets the ispC gene product, DXR. This selection is especially challenging since it is a relatively weak in E. coli, with multiple naturally occurring pathways for resistance. We identified several previously unreported mutations that confer fosmidomycin resistance, in highly conserved sites that also exist in pathogens including the malaria-inducing Plasmodium falciparum. This approach may have implications for the isolation of resistance-conferring mutations and may affect the design of future generations of fosmidomycin-based drugs. |
Databáze: | OpenAIRE |
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