ÃÞ »¨ ѧ ±¨   Cotton Science  2005£¬17(6)£º323-327

 

Effect of Two Methods of Protein Extraction from Cotton Ovule and Fiber
LIU Kang£¬HU Feng-ping£¬ZHANG Tian-zhen
(Cotton Institute£¬Nanjing Agricultural University£¬State Key Laboratory of Crop Genetics and Germplasm Enhancement£¬Nanjing Agricultural University£¬Nanjing 210095£¬China )ª¤ª¤

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¤
Key words£ºcotton; ovule; fiber; proteinª¤ª¤ [Full Text, 6052KB]