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[求助]
求助:纖維素合成酶 酶活性測定方法,謝謝!
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| 求助,急需纖維素合成酶酶活性的測定方法,謝謝!第一次發(fā)帖,不對的地方見諒!謝謝! |
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纖維素合成酶活性測定比較困難,常見的方法如下 Cellulose synthase activity in membrane preparations of A. xylinum was determined according to the procedure of Lin and Brown (13). For enzyme assays, approximately 150 mg of membrane protein was incubated at 308C for 30 min in a reaction mixture with a final concentration of 20 mM UDP-[14C]glucose and 20 mM cyclic diguanylic acid (c-di-GMP) (gift from J. H. van Boom, Department of Organic Chemistry, Leiden, The Netherlands), and the radioactivity present in the alkali-insoluble product obtained after boiling was used to determine the cellulose synthase activity. Protein concentrations were determined by Peterson’s modification of the micro-Lowry method (19) with a protein assay kit (Sigma). 方法參考自 Identification of a second cellulose synthase gene (acsAII) in Acetobacter xylinum J. Bacteriol. 1995, 177(18):5276. 涉及到的其他文獻包括 13 Lin, F. C., and R. M. Brown, Jr. 1989. Purification of cellulose synthase from Acetobacter xylinum, p. 473–492. In C. Schuerch (ed.), Cellulose and wood— chemistry and technology. John Wiley and Sons, New York. 19 Peterson, G. L. 1977. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal. Biochem. 83:346–356. |

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