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首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >A simple protocol for protein extraction of recalcitrant fruit tissues suitable for 2-DE and MS analysis
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A simple protocol for protein extraction of recalcitrant fruit tissues suitable for 2-DE and MS analysis

机译：适用于2-DE和MS分析的顽固性水果组织蛋白提取的简单协议

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摘要

Fruit tissues are considered recalcitrant plant tissue for proteomic analysis. Three phenol-free protein extraction procedures for 2-DE were compared and evaluated on apple fruit proteins. Incorporation of hot SDS buffer, extraction with TCA/acetone precipitation was found to be the most effective protocol. The results from SDS-PAGE and 2-DE analysis showed high quality proteins. More than 500 apple polypeptides were separated on a small scale 2-DE gel. The successful protocol was further tested on banana fruit, in which 504 and 386 proteins were detected in peel and flesh tissues, respectively. To demonstrate the quality of the extracted proteins, several protein spots from apple and banana peels were cut from 2-DE gels, analyzed by MS and have been tentatively identified. The protocol described in this study is a simple procedure which could be routinely used in proteomic studies of many types of recalcitrant fruit tissues.
机译：水果组织被认为是蛋白质组学分析的顽强植物组织。比较了三种针对苹果酸的无酚蛋白质提取方法，并对苹果果实蛋白质进行了评估。发现加入热的SDS缓冲液，用TCA /丙酮沉淀萃取是最有效的方法。 SDS-PAGE和2-DE分析的结果显示了高质量的蛋白质。在小型2-DE凝胶上分离了500多个苹果多肽。在香蕉果实上进一步测试了成功的方案，在果皮和果肉组织中分别检测到504和386蛋白。为了证明提取蛋白质的质量，从2-DE凝胶上切下了苹果和香蕉皮中的几个蛋白质斑点，通过MS进行了分析，并已初步鉴定。这项研究中描述的协议是一个简单的过程，可以常规用于许多类型的顽固性水果组织的蛋白质组学研究。

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