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Optimization of protein extraction and proteomic studies in Cenchrus polystachion (L.) Schult

机译:Cenchrus polystachion(L.)Schult中蛋白质提取和蛋白质组学研究的优化

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摘要

Apomicts have been studied at their genetic levels, but there are no any direct evidence of its mechanism. In order to understand the mechanism involved, a close relative of , , an apomictic species was explored for more insights into protein expression in reproductive structures. Optimization of protein extraction was studied with the leaf tissue and optimized protocol was extrapolated to other five tissues. The phenol-based protein extraction emerged as the best method for plant leaf tissue providing a better protein yield, separation of bands, removal of non-protein components like polyphenolic compounds and nucleic acids. The proteome analysis of leaf, stigma, immature ovary, seed, anther sac and pollen tissues of were carried out identifying a total of 135407 proteins against the Poaceae database from UNIPROT/TrEMBL. The target candidate proteins found in all the tissues were identified and mainly comprised of Actin Protein, PIP, Starch Synthase, ATP Synthase, Glutathione S Transferase, Dehydroascorbate reductase, Ascorbate peroxidase and heat shock proteins. Visualization and descriptive statistics conveyed all the necessary information to understand the differential expression of proteins in . This study forms a base to understand the role of tissue specific expressed proteins in an apomictic plant.
机译:已经在其遗传水平上研究了无融合生殖,但是没有任何直接证据证明其机制。为了了解所涉及的机制,探索了无融合生殖物种的近亲,以深入了解生殖结构中的蛋白质表达。用叶组织研究了蛋白质提取的优化,并将优化的方案外推至其他五种组织。基于酚的蛋白质提取方法已成为植物叶片组织的最佳方法,可提供更好的蛋白质产量,分离条带,去除非蛋白质成分(如多酚化合物和核酸)。对叶,柱头,未成熟的卵巢,种子,花药囊和花粉组织的蛋白质组学分析,从UNIPROT / TrEMBL的禾本科数据库中鉴定出总共135407种蛋白质。鉴定了在所有组织中发现的目标候选蛋白,主要由肌动蛋白,PIP,淀粉合酶,ATP合酶,谷胱甘肽S转移酶,脱氢抗坏血酸还原酶,抗坏血酸过氧化物酶和热休克蛋白组成。可视化和描述统计传达了所有必要的信息,以了解蛋白质在大肠杆菌中的差异表达。该研究为了解组织特异性表达蛋白在无融合生殖植物中的作用奠定了基础。

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