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水稻花药总蛋白质双向电泳方法的改良与优化(简报)
IMPROVEMENT OF TWO-DIMENSIONAL GEL ELECTROPHORESIS OF TOTAL PROTEINS FROM RICE ANTHERS
【摘要】 <正>双向电泳作为蛋白质组学核心技术之一,目前已广泛地应用在植物领域,并且成功应用于水稻代谢和调节等方面的研究。谢锦云等利用溶液法提取温敏核不育水稻花药总蛋白质,利用pH3-10线性胶条分离,经银染显色后检测到约1,000个
【Abstract】 This paper reported an improvement in 2-D gel electrophoresis of the proteome in Honglian cytoplasmic male sterile rice. An IPGphor?unit with immobile pH gradient strips was used as the first dimension and SDS-PAGE as the second. The total anther proteins were extracted using TCA/acetone and then were washed 5-6 times with acetone till the proteins were white and clean,and then tributylphosphine and DTT were added into the rehydration buffer to improve the solubility of the proteins. The 2-D gel was stained by both methods of coomassie blue G-250 and silver. Extraction of proteins,pH of the strips and rehydration of the strips were optimized and compared. Higher repeatability and better separating protein pattern could be gained by this technique.
- 【文献出处】 分子细胞生物学报 ,Journal of Molecular Cell Biology , 编辑部邮箱 ,2006年05期
- 【分类号】S511
- 【被引频次】17
- 【下载频次】320