Plant Science


    Protocols in Current Issue
    Chromatin Immunoprecipitation (ChIP) to Assess Histone Marks in Auxin-treated Arabidopsis thaliana Inflorescence Tissue
    Authors:  André Kuhn and Lars Østergaard, date: 12/05/2020, view: 885, Q&A: 0
    [Abstract] Chromatin immunoprecipitation coupled with quantitative PCR (ChIP-qPCR) or high-throughput sequencing (ChIP-seq) has become the gold standard for the identification of binding sites of DNA binding proteins and the localization of histone modification on a locus-specific or genome-wide scale, respectively. ChIP experiments can be divided into seven ...
    Ratiometric Measurement of Protein Abundance after Transient Expression of a Transgene in Nicotiana benthamiana
    Authors:  Aashima Khosla and David C. Nelson, date: 09/05/2020, view: 1876, Q&A: 0
    [Abstract] Ratiometric reporters are tools to dynamically measure the relative abundance of a protein of interest. In these systems, a target protein fused to a fluorescent or bioluminescent reporter is expressed with fixed stoichiometry to a reference protein fused to a second reporter. Both fusion proteins are encoded on a single transcript but are ...
    High-resolution and Deep Phylogenetic Reconstruction of Ancestral States from Large Transcriptomic Data Sets
    Authors:  Sumanth Kumar Mutte and Dolf Weijers, date: 03/20/2020, view: 1939, Q&A: 0
    [Abstract] Phylogenetics is an important area of evolutionary biology that helps to understand the origin and divergence of genes, genomes and species. Building meaningful phylogenetic trees is needed for the accurate reconstruction of the past. To achieve a correct phylogenetic understanding of genes or proteins, reliable and robust methods are needed to ...
    Measuring Protein Half-life in Arabidopsis thaliana
    Authors:  Hongqing Guo and Yanhai Yin, date: 08/05/2019, view: 2304, Q&A: 0
    [Abstract] Post-translational modifications play important roles in controlling protein function and can lead to altered protein stability. Protein stability can be determined after treatment with the protein synthesis inhibitor Cycloheximide. Cycloheximide is a translational inhibitor that inhibits protein synthesis via cytoplasmic ribosomes. Here we ...
    Extraction and Purification of Laccases from Rice Stems
    Authors:  Chenna Swetha and P. V. Shivaprasad, date: 04/05/2019, view: 3190, Q&A: 0
    [Abstract] Laccases are found in cell walls of plants in very low amounts. This protocol provides an efficient method to purify laccases from rice stems. The method involves three steps: 1) Isolation of total protein from rice stems using buffers with high salt concentration to extract protein from cell walls; 2) Purification of laccases using concanavalin-A ...
    Using Arabidopsis Mesophyll Protoplasts to Study Unfolded Protein Response Signaling
    Authors:  Yan Bao and Diane C. Bassham, date: 12/05/2018, view: 3476, Q&A: 0
    [Abstract] Various environmental stresses or artificial reagents can trigger unfolded protein accumulation in the endoplasmic reticulum (ER) due to the folding capacity of the ER being exceeded. This is termed ER stress, and triggers the unfolded protein response (UPR). Assays for activation of the UPR in plants include Tunicamycin (Tm)- or dithiothreitol ...
    Separation of Thylakoid Protein Complexes with Two-dimensional Native-PAGE
    Authors:  Marjaana Rantala, Virpi Paakkarinen and Eva-Mari Aro, date: 07/05/2018, view: 5666, Q&A: 0
    [Abstract] The hierarchical composition and interactions of the labile thylakoid protein complexes can be assessed by sequential 2D-native gel-electrophoresis system. Mild non-ionic detergent digitonin is used to solubilize labile protein super-and megacomplexes, which are then separated with first-dimension blue native polyacrylamide gel electrophoresis ...
    Detecting the Interaction of Double-stranded RNA Binding Protein, Viral Protein and Primary miRNA Transcript by Co-immunoprecipitation in planta
    Authors:  Lijia Zheng, Chao Zhang, Jianguo Wu and Yi Li, date: 05/05/2018, view: 3522, Q&A: 0
    [Abstract] MicroRNAs (miRNAs) play important roles in plant growth, development, and response to infection by microbes. Double-stranded RNA binding protein 1 (DRB1) facilitates the processing of primary miRNA transcripts into mature miRNAs. Recently, we found that NS3 protein encoded by rice stripe virus (RSV) associates with DRB1 and promotes miRNA ...
    Boron Uptake Assay in Xenopus laevis Oocytes
    Authors:  Sheliang Wang, Namiki Mitani-Ueno and Junpei Takano, date: 03/05/2018, view: 4990, Q&A: 0
    [Abstract] Boron (B) is essential for plant growth and taken up by plant roots as boric acid. Under B limitation, B uptake and translocation in plants are dependent on the boric acid channels located in the plasma membrane. Xenopus leavis oocyte is a reliable heterologous expression system to characterize transport activities of boric acid channels ...
    Ribosomal RNA N-glycosylase Activity Assay of Ribosome-inactivating Proteins
    Authors:  Rosario Iglesias, Lucía Citores and José M. Ferreras, date: 03/20/2017, view: 8072, Q&A: 0
    [Abstract] Ribosome-inactivating proteins (RIPs) are enzymes that irreversibly inactivate ribosomes as a consequence of their N-glycosylase (EC activity. The enzyme cleaves the N-glycosidic bond between the adenine No. 4324 from the 28S rRNA and its ribose in rat ribosomes (or the equivalent adenine in sensitive ribosomes from other organisms). ...

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