NR | Nitrate reductase, assimilatory
AS08 310 | Clonality: Polyclonal | Host: Rabbit | Reactivity: A. thaliana, H. vulgare, C. reinhardtii, red alga Gracilaria gracilis, Medicago sativa, diatom Thalassiosira sp., P. tricornutum Bohlin, P. yunnanensis Dode, P. notoginseng, S. lycopersicum, S. tuberosum
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103 kDa | 117 kDa
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Aspergilus niger, Emiliania huxleyi, Tisochrysis lutea
Reactant: Chlamydomonas reinhardtii (Green Alga)
Application: Western Blotting
Pudmed ID: 25653846
Figure Number: 1A
Published Date: 2015-02-06
First Author: Johnson, E. A. & Lecomte, J. T.
Impact Factor: NoneOpen Publication
Purification of THB1 fromC. reinhardtii cell culture.Protein samples from different stages of purification were analyzed by electrophoresis. (A) Samples of protein extracts before and after lysis ofC. reinhardtii cells with liquid nitrogen. Proteins separated on 16.5% Tris-tricine gel then transferred to nitrocellulose and immunostained with antibodies against proteins localized to different cellular compartments. Lysis by liquid nitrogen enriches the lysate with soluble proteins without enrichment of proteins found in major algal organelles. Histone H3 protein is located in the nucleus, nitrate reductase is a soluble cytosolic protein and the ATP synthase ? subunit is part of the thylakoid membrane of the chloroplast. (B) Samples from different steps in the purification procedure were separated by electrophoresis. Following separation, the proteins within the gel were visualized using silver stain. (C) Proteins prepared identically to those detected in panel B were transferred to nitrocellulose followed by immunostaining with polyclonal antibodies specific for THB1. In addition to the protein samples, a lane was used for molecular weight markers (Spectra LR, ThermoScientific). The numbers indicated between the panels represents location of the markers (kDa molecular mass). WC, whole cell protein extract. Lys, protein extract lysate, and Pel, protein pellet following liquid nitrogen fracturing. QFF, concentrated sample following anion exchange chromatography. S75, concentrated sample following separation on the Superdex 75 column.
Application: Western Blotting
Pudmed ID: 33418923
Journal: Int J Mol Sci
Figure Number: 4A
Published Date: 2021-01-06
First Author: Kim, J., Chang, K. S., et al.
Impact Factor: 5.542Open Publication
Expression of NR and APT in edited mutants compared with the WT. (A) Western blot analysis with a specific antibody against nitrate reductase (upper panel). Total protein content was assessed by Coomassie staining (lower panel). ATP-? was used as a reference protein. (B) Comparison of gene and amino acid sequence of APT between edited mutants and the WT. PCR amplification of APT from cDNA of apt mutants and WT cells (upper panel) and corresponding sequences (lower panels), showing the presence of frameshift mutations. M: size marker.
Using this antibody genome editing in Chlorella vulgaris UTEX395 by CRISPR-Cas9 system has been demonstrated as described in Kim et al. (2021)
Chemiluminescent detection is advised for NR detection using this antibody.
Assimilatory nitrate reductase (NR), (EC.18.104.22.168) catalyses the reduction of nitrate to nitrite in the cytoplasm. Plants contain 2 forms of NR: NADH-NR (most common form in plants and algae, predominantly found in green tissues) and NAD(P)H-NR (uses NADH or NADPH as the electron donor, constitutively expressed in plants at a low level). NADH-NR is a homodimer of two identical subunits (100-115 kDa each, hold together by a Mo-cofactor) each of them coded by up to three genes (NR1-3, NIA1-NIA3).
Kim et al. (2021). Establishment of a Genome Editing Tool Using CRISPR-Cas9 in Chlorella vulgaris UTEX395. Int J Mol Sci. 2021 Jan 6;22(2):E480. doi: 10.3390/ijms22020480. PMID: 33418923.
Prinsi et al. (2021). Biochemical and Proteomic Changes in the Roots of M4 Grapevine Rootstock in Response to Nitrate Availability. Plants 10, no. 4: 792. https://doi.org/10.3390/plants10040792
Maresca et al. (2021) Biological responses to heavy metal stress in the moss Leptodictyum riparium (Hedw.) Warnst. Ecotoxicol Environ Saf. 2022 Jan 1;229:113078. doi: 10.1016/j.ecoenv.2021.113078. Epub 2021 Dec 17. PMID: 34929502.
Zhang et al. (2020). Hydrogen sulfide and rhizobia synergistically regulate nitrogen (N) assimilation and remobilization during N deficiency-induced senescence in soybean. Plant Cell Environ. 2020 Feb 3. doi: 10.1111/pce.13736.
Dongxu et al. (2020). Magnesium reduces cadmium accumulation by decreasing the nitrate reductase-mediated nitric oxide production in Panax notoginseng roots. Journal of Plant Physiology. Available online 7 February 2020, 153131
Jayawardena et al. (2016). Elevated CO2 plus chronic warming reduces nitrogen uptake and levels or activities of nitrogen -uptake and -assimilatory proteins in tomato roots. Physiol Plant. 2016 Nov 28. doi: 10.1111/ppl.12532. [Epub ahead of print]
Chen et al. (2016). The role of nitric oxide signalling in response to salt stress in Chlamydomonas reinhardtii. Planta. 2016 Sep;244(3):651-69. doi: 10.1007/s00425-016-2528-0. Epub 2016 Apr 26.
Cheng et al. (2015). Quantitative proteomics analysis reveals that S-nitrosoglutathione reductase (GSNOR) and nitric oxide signaling enhance poplar defense against chilling stress. Planta. 2015 Aug 2.
Zhang et al. (2014). Heterologous expression of AtPAP2 in transgenic potato influences carbon metabolism and tuber development. FEBS Lett. 2014 Aug 27. pii: S0014-5793(14)00621-8. doi: 10.1016/j.febslet.2014.08.019.
Beyzaei et al. (2014). Response of Nitrate Reductase to Exogenous Application of 5-Aminolevulinic Acid in Barley Plants. J. Plant Growth Regulation, April 2014.
Frada et al. (2013). Quantum requirements for growth and fatty acid biosynthesis in the marine diatom Phaeodactylum tricornutum (Bacilloriophyceae) in nitrogen replete and limited conditions. J. Phycology. Diatom growth and lipid efficiency
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