Controlling the Expression of Rhizobial Genes During Nodule Development with Elements and An Inducer of the lac Operon
| Autor: | Jodie M. Box, K. Dale Noel |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Isopropyl Thiogalactoside
Time Factors Transcription Genetic Physiology Operon Repressor lac operon Lac repressor Biology Plant Root Nodulation Microbiology Bacterial Proteins Gene expression Escherichia coli Lac Repressors Inducer Promoter Regions Genetic Symbiosis Glucuronidase Sequence Deletion Phaseolus Regulation of gene expression Escherichia coli Proteins O Antigens food and beverages Gene Expression Regulation Bacterial General Medicine biology.organism_classification carbohydrates (lipids) Lac Operon Genes Bacterial Rhizobium Agronomy and Crop Science Medicago sativa Plasmids Sinorhizobium meliloti |
| Zdroj: | Molecular Plant-Microbe Interactions®. 24:478-486 |
| ISSN: | 1943-7706 0894-0282 |
| DOI: | 10.1094/mpmi-07-10-0155 |
| Popis: | A simple strategy was tested for imposing artificial regulation of rhizobial genes during nodule development. Isopropyl-β-d-1-thiogalactoside (IPTG) was added to liquid root media to sustain expression of rhizobial genes controlled by Escherichia coli lac promoter/operators and repressor gene lacI. Conversely, a rinsing protocol was devised to remove IPTG sufficiently that genes could be repressed after having been induced. gusA under this control exhibited clearly delineated expression and repression in both the determinate Rhizobium etli–Phaseolus vulgaris and the indeterminate Sinorhizobium meliloti–Medicago sativa symbioses. Apparently, IPTG was taken up in sufficiently undegraded concentrations that gene expression was derepressed even in interior portions of the nodule. Moreover, the rinsing protocol led to obvious repression of gusA. Importantly, no deleterious effects of IPTG on nodule development, infection, or nitrogen fixation were observed. An R. etli CE3 gene required for lipopolysaccharide O antigen and infection on bean was put under this control by means of a two-plasmid construct. When this construct was added to a strain with a null mutation in this gene, infection, nodule development, and nitrogenase activity all depended on the length of time before IPTG was rinsed from the roots after inoculation. |
| Databáze: | OpenAIRE |
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