In vitro assembly of plant tubulin in the absence of microtubule-stabilizing reagents |
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Authors: | Shanjin Huang Haiyun Ren Ming Yuan |
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Affiliation: | (1) Key Laboratory of Plant Physiology and Biochemistry of Ministry of Agriculture, Department of Plant Sciences, College of Biological Sciences, China Agricultural University, 100094 Beijing, China;(2) Key Laboratory of Cell Proliferation and Regulation Biology of Ministry of Education, College of Life Sciences, Beijing Normal University, 100875 Beijing, China |
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Abstract: | The assembly of microtubules is essential for physiological functions of microtubules. Addition of microtubule-stabilizing reagents or microtubule “seeds” is usually necessary for plant tubulin assemblyin vitro, which hinders the investigation of plant microtubule dynamics. In the present note, highly purified plant tubulins have been obtained from lily pollen, a non-microtubule-stabilizing reagent or microtubule “seed” system for plant tubulin assembly has been established and the analysis of plant tubulin assembly performed. Experiment results showed that purified tubulin polymerizedin vitro, and a typical microtubule structure was observed with electron microscopy. The kinetics curve of tubulin assembly exhibited typical “parabola”. The presence of taxol significantly altered the character of plant tubulin assembly, including that abnormal microtubules were assembled and the critical concentration for plant tubulin assembly was decreased exceedingly from 3 mg/mL in the absence of taxol to 0.043 mg/mL in ihe presence of taxol. |
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Keywords: | lily pollen tubulin microtubule tubulin assembly |
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