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ÃÞ »¨ ѧ ±¨ Cotton Science 2005£¬17(6)£º323-327
Effect of Two Methods of Protein Extraction from Cotton Ovule and Fiber Abstract£º Total protein of cotton ovule and fiber were extracted using Trichloroacetic acid (TCA)-acetone precipitation method and phenol isolation method£¬respectively£¬we found that for the TCA-acetone precipitation method£¬the protein harvest rate was 2.3 mg¡¤g-1. The protein sample was separated with 2-dimension gel electrophoresis (2-DE) and visualized with colloidal Coomassie Brilliant Blue G-250 (cCBB G-250) and silver staining methods respectively£¬in cCBB G-250 gels£¬few protein spots were detected£¬no considerable change observed in silver stained gel as well. For phenol isolation method£¬however£¬the harvest rate reached 10.4 mg¡¤g-1£¬which was as 5 folds high as that of TCA-acetone precipitation method£¬923 spots were detected on cCBB stained 2-DE gel£¬ and 1061 spots detected on silver stained 2-DE gel£¬ the isoelectric poit (pI) values of the proteins ranged from 3 to 10£¬focusing between 4.3 and 8.6. The results demonstrated that phenol isolation method£¬which may harvest more kinds of protein with wider pI distribution range and better dissolvability£¬is easy-handling and thus is optimal for extracting cotton ovule and fiber protein in proteomic research.ª¤ |
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