Tissue and Cell-type Localization and Partial Characterization of a Xylem Papain-type Cysteine Protease from Arabidopsis
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چکیده
............................................................................................................................... ii Acknowledgements .............................................................................................................. iii List of Figures ...................................................................................................................... vii Chapter 1: Literature Review ............................................................................................ 1 Chapter 2: Materials and Methods ................................................................................... 9 2.1 Expression in E. coli and purification of XCP1 for antibody production..... 9 2.1.1 Construction of polyhistidine-tagged XCP1 ........................................ 9 2.1.2 Preparation of chemically competent E. coli ....................................... 9 2.1.3 Transformation of competent M15 cells.............................................. 11 2.1.4 Analysis of transformants..................................................................... 11 2.1.5 Expression of XCP1H6 and purification by metal-chelate chromatography............................................................................................. 12 2.1.6 Activation of protease .......................................................................... 13 2.1.7 Protease activity gel ............................................................................. 14 2.2 Developmental time course and tissue-level localization of XCP1 protein in the root-hypocotyl of Arabidopsis ...................................................................... 14 2.2.1 Plant growth and separation of xylem and bark................................... 15 2.2.2 Preparation of tissue extracts for immunoblot analysis ....................... 15 2.2.3 Production and affinity purification of anti-XCP1 antibody................ 15 2.2.4 Immunoblot analysis ............................................................................ 16
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تاریخ انتشار 2000