| Phosphinothricin resistance in Aspergillus niger and its utility as a selectable transformation marker. | |
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MedLine Citation:
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PMID: 18479949 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Aspergillus niger is moderately susceptible to inhibition by phosphinothricin (PPT)-a potent inhibitor of glutamine synthetase. This growth inhibition was relieved by L-glutamine. PPT inhibited A. niger glutamine synthetase in vitro (K(I), 54 microM) and the inhibition was competitive with L-glutamate. The bar gene, imparting resistance to PPT, was successfully exploited as a dominant marker to transform this fungus. Very high PPT concentrations were required in the overlay for selection. Apart from bar transformants, colonies spontaneously resistant to PPT were frequently encountered on selection media. Reasons for such spontaneous resistance, albeit of moderate growth phenotype, were sought using one such isolate (SRPPT). The SRPPT isolate showed a 2-3-fold decrease in its glutamate uptake rate. Elevated external glutamate levels further suppressed the PPT-induced growth inhibition. Cellular entry of PPT could be through the L-glutamate uptake system thereby accounting for the observed spontaneous resistant phenotype. These results were useful in the fine-tuning of bar-selection in A. niger. |
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Authors:
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Manmeet Ahuja; Narayan S Punekar |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2008-04-08 |
Journal Detail:
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Title: Fungal genetics and biology : FG & B Volume: 45 ISSN: 1096-0937 ISO Abbreviation: Fungal Genet. Biol. Publication Date: 2008 Jul |
Date Detail:
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Created Date: 2008-06-09 Completed Date: 2008-07-01 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9607601 Medline TA: Fungal Genet Biol Country: United States |
Other Details:
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Languages: eng Pagination: 1103-10 Citation Subset: IM |
Affiliation:
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Biotechnology Group, School of Bioscience and Bioengineering, Indian Institute of Technology Bombay, Mumbai 400076, India. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Aminobutyric Acids
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metabolism,
pharmacology* Aspergillus niger / drug effects*, enzymology, genetics*, metabolism Drug Resistance, Fungal* Enzyme Inhibitors / metabolism, pharmacology Fungal Proteins / antagonists & inhibitors, metabolism Genetic Markers Glutamate-Ammonia Ligase / antagonists & inhibitors, metabolism Glutamic Acid / metabolism Kinetics Transformation, Genetic* |
| Chemical | |
Reg. No./Substance:
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0/Aminobutyric Acids; 0/Enzyme Inhibitors; 0/Fungal Proteins; 0/Genetic Markers; 51276-47-2/phosphinothricin; 56-86-0/Glutamic Acid; EC 6.3.1.2/Glutamate-Ammonia Ligase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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