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选用干旱敏感型栽培烟草品种中烟100旺长后期的叶片和感黑胫病栽培烟草品种小黄金1025苗期的叶片作为实验材料,总蛋白质提取分别采用稍加改进的TCA/丙酮法和由蛋白质分级提取、酚抽提及丙酮洗涤相结合的方法。对中烟100旺长后期烟草叶片采用的总蛋白质TCA/丙酮提取方法进行了优化、对其使用的IPG胶条pH范围做了初步筛选,并在此研究基础上,对采用第二种提取方法获得的小黄金1025苗期烟草叶片总蛋白质样品进行了不同上样量对双向电泳结果影响的比较实验。初步建立起适合烟草叶片蛋白质组研究的双向电泳实验体系。
The leaves of the susceptible cultivar Zhongyan 100 Wangchang and the susceptible black shank cultivars were used as the experimental materials. The total protein was extracted by TCA / acetone method with slight improvement and protein by protein Grading extraction, phenol extraction and acetone washing method. The total protein TCA / acetone extraction method was optimized for the late tobacco leaf during 100 Mongolian-long tobacco cultivation. The pH range of the used IPG adhesive tape was preliminarily screened. On the basis of this study, the second extraction method The obtained small gold 1025 seedling tobacco leaf total protein samples were subjected to different loading amount of two-dimensional electrophoresis results of comparative experiments. Preliminary establishment of two-dimensional electrophoresis experiment system suitable for tobacco leaf proteome research.