Arabidopsis
| العنوان: | Arabidopsis |
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| المؤلفون: | Abramov, Aleksej, Hoffmann, Thomas, Stark, Timo D., Zheng, Linlin, Lenk, Stefan, Hammerl, Richard, Lanzl, Tobias, Dawid, Corinna, Schon, Chris-Carolin, Schwab, Wilfried, Gierl, Alfons, Frey, Monika |
| المصدر: | Engineering of benzoxazinoid biosynthesis in Arabidopsis thaliana: Metabolic and physiological challenges, pp. 1-15 in Phytochemistry (112947) 192 11 |
| سنة النشر: | 2021 |
| المجموعة: | Zenodo |
| مصطلحات موضوعية: | Biodiversity, Taxonomy, Plantae, Tracheophyta, Magnoliopsida, Brassicales, Brassicaceae, Arabidopsis |
| الوصف: | 5.9. Isolation of Arabidopsis microsomes and P450 assay 15 g of rosette leaves of 28 dag Arabidopsis plants were ground in 200 mL extraction buffer (100 mM ascorbic acid, 1 mM EDTA, 100 mM Tris, 20% v/v glycerol, 20% w/v Sucrose, 5 mM Dithiothreitol) with 4.5 g Polyklar AT (Merck) and sea sand. The raw extract was filtered through cloth and centrifuged twice at 15 000 g for 10 min. Microsomes were isolated from the supernatant by centrifugation at 120 000 g for 40 min and resuspended in 1 mL suspension buffer (50 mM potassium phosphate buffer pH 7.5, 20% v/v Glycerol, 1 mM Dithiothreitol). The integrity of the microsomes was tested by measuring the cytochrome C reductase activity as described by Urban et al. (1990). The in vitro activity of the BX P450 enzymes was tested by incubation of 1 mg total microsomal protein with the respective substrate (2 mM Indole, 1 mM ION, 250 μM HION, 200 μM HBOA) in 100 mM potassium phosphate buffer pH 7.5 and 1 mM NADPH at room temperature. The reaction was stopped after 30 min by addition of 1 vol methanol and precipitated protein was pelleted by centrifugation. 2.5 vol of 100 mM acetic acid were added to the supernatant and the products were extracted three times with 2 vol of ethyl acetate. The solvent was evaporated in a vacuum centrifuge, the remaining products were resolved in methanol and analysed by HPLC. ; Published as part of Abramov, Aleksej, Hoffmann, Thomas, Stark, Timo D., Zheng, Linlin, Lenk, Stefan, Hammerl, Richard, Lanzl, Tobias, Dawid, Corinna, Schon, Chris-Carolin, Schwab, Wilfried, Gierl, Alfons & Frey, Monika, 2021, Engineering of benzoxazinoid biosynthesis in Arabidopsis thaliana: Metabolic and physiological challenges, pp. 1-15 in Phytochemistry (112947) (112947) 192 on page 11, DOI:10.1016/j.phytochem.2021.112947 |
| نوع الوثيقة: | other/unknown material |
| اللغة: | unknown |
| العلاقة: | info:eu-repo/semantics/altIdentifier/url/http://treatment.plazi.org/id/5B752120683AFFF3FFAFFB52FDD3687ETest; http://publication.plazi.org/id/A74C59586830FFF9FFCBFFEBFFDE6F68Test; https://sibils.text-analytics.ch/search/collections/plazi/5B752120683AFFF3FFAFFB52FDD3687ETest; https://zenodo.org/communities/biosyslitTest; https://zenodo.org/record/8270283Test; https://doi.org/10.5281/zenodo.8270283Test; oai:zenodo.org:8270283 |
| DOI: | 10.5281/zenodo.8270283 |
| الإتاحة: | https://doi.org/10.5281/zenodo.8270283Test https://doi.org/10.1016/j.phytochem.2021.112947Test https://doi.org/10.5281/zenodo.8270282Test https://zenodo.org/record/8270283Test |
| حقوق: | info:eu-repo/semantics/openAccess ; https://creativecommons.org/publicdomain/zero/1.0/legalcodeTest |
| رقم الانضمام: | edsbas.EEE73CB5 |
| قاعدة البيانات: | BASE |
| DOI: | 10.5281/zenodo.8270283 |
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