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    Protocols in Current Issue
    Measuring Secretion of Capsidiol in Leaf Tissues of Nicotiana benthamiana
    Authors:  Teruhiko Kuroyanagi, Maurizio Camagna and Daigo Takemoto, date: 08/05/2018, view: 1263, Q&A: 0
    [Abstract] Plant species produce a wide variety of antimicrobial metabolites to protect themselves against potential pathogens in natural environments. Phytoalexins are low molecular weight compounds produced by plants in response to attempted attacks of pathogens. Accumulation of phytoalexins in attacked plant tissues can inhibit the growth of penetrating ...
    Histone Deubiquitination Assay in Nicotiana benthamiana
    Authors:  Shujing Liu and Lars Hennig, date: 03/05/2018, view: 1486, Q&A: 0
    [Abstract] Histone modifications are a group of post-translational modifications on histones which can alter chromatin structure and affect gene expression. Histone ubiquitination is a histone modification found in particular on histone H2A and H2B. Histone ubiquitination can be reversed by ubiquitin-specific proteases (UBP). Here, we describe an in vivo ...
    Transient Gene Expression for the Characteristic Signal Sequences and the Estimation of the Localization of Target Protein in Plant Cell
    [Abstract] We have proposed and tested a method for characterization of the signal sequences and determinations of target protein localization in a plant cell. This method, called the AgI-PrI, implies extraction of protoplasts from plant tissues after agroinfiltration. The suggested approach combines the advantages of two widely used methods for transient ...
    Use of Geminivirus for Delivery of CRISPR/Cas9 Components to Tobacco by Agro-infiltration
    Authors:  Kangquan Yin, Ting Han and Yule Liu, date: 04/05/2017, view: 6531, Q&A: 0
    [Abstract] CRISPR/Cas9 system is a recently developed genome editing tool, and its power has been demonstrated in many organisms, including some plant species (Wang et al., 2016). In eukaryotes, the Cas9/gRNA complexes target genome sites specifically and cleave them to produce double-strand breaks (DSBs), which can be repaired by non-homologous end ...
    Pot Level Drought Stress Tolerance Assay in Tobacco through Plant Phenotyping and Antioxidant Assay
    Authors:  Priyanka Das, Nita Lakra, Kamlesh Kant Nutan, Sneh L Singla-Pareek and Ashwani Pareek, date: 10/05/2015, view: 7049, Q&A: 0
    [Abstract] Drought is an important abiotic factor which has a huge detrimental impact on crop productivity. Study of plant responses towards drought stress and investigating the mechanism of drought tolerance is crucial for achieving the target of developing drought-tolerant plants. Phenotyping is a cost effective approach which can be adopted to evaluate ...
    In vitro Reconstitution Assay of miRNA Biogenesis by Arabidopsis DCL1
    Authors:  Tian Wang, Claudia Castillo-González, Lan You, Rui Li, Liwei Wen, Hongliang Zhu and Xiuren Zhang, date: 04/20/2015, view: 5328, Q&A: 0
    [Abstract] microRNAs (miRNAs) are small non-coding RNAs, regulating most if not all, biological processes in eukaryotic organisms. miRNAs are initially processed from primary transcripts (pri-miRNAs) to produce miRNA precursors (pre-miRNAs), that are further processed into miRNA and its complementary strands (miRNA/*). In Arabidopsis, and possibly other ...
    Phosphatase Protection Assay: 14-3-3 Binding Protects the Phosphate group of RSG from λ Protein Phosphatase
    Authors:  Takeshi Ito and Yohsuke Takahashi, date: 02/05/2015, view: 6111, Q&A: 0
    [Abstract] 14-3-3 proteins regulate diverse cellular processes in eukaryotes by binding to phospho-serine or threonine of target proteins. One of the physiological functions of 14-3-3 is to bind and protect phosphate groups of the target proteins against phosphatases. REPRESSION OF SHOOT GROWTH (RSG) is a tobacco (Nicotiana tabacum) transcription ...
    Cytokinin Analysis: Sample Preparation and Quantification
    Authors:  Martin Schäfer, Michael Reichelt, Ian T. Baldwin and Stefan Meldau, date: 07/05/2014, view: 9223, Q&A: 0
    [Abstract] Cytokinins are a group of phytohormones discovered about half a decade ago by Miller et al. (1955) and Skoog et al. (1965). Since then they were found to participate in many plant physiological processes, including the regulation of the source/sink transitions, plant growth and organ development, responses to environmental ...
    Preparation of Arabinogalactan Glycoproteins from Plant Tissue
    Author:  Derek T. A. Lamport, date: 10/05/2013, view: 7174, Q&A: 1
    [Abstract] This supplements an earlier protocol (Popper, 2011) for the extraction and assay of cell surface arabinogalactan proteins (AGPs). These highly glycosylated glycoproteins (~95% carbohydrate) contain numerous glycomodules with paired glucuronic acid residues that bind Ca2+ in a pH dependent manner (Lamport and Varnai, 2013). Classical ...
    Reverse Zymmogram Analysis for the Detection of Protease Inhibitor Activity
    Authors:  Lilia Bernal, Florencia García-Campusano, Edgar Nájera and Felipe Cruz-García, date: 08/20/2013, view: 7706, Q&A: 0
    [Abstract] This protocol describes a gel-based procedure to detect protease inhibitor activity. In this method gelatin is used as a substrate for proteolysis and is copolymerized within the polyacrylamide matrix. Protein extracts are fractioned by SDS-PAGE and then the gel is treated with the protease of interest, which degrades gelatin, except in the areas ...



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