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Author:
Date:
2015-02-20
[Abstract] Plant transformation and exogenous protein expression is essential for molecular biology and biotechnology. Current approaches of stable plant transformation might be problematic and very time-consuming. Because of this, transient expression in protoplasts has become valuable alternative, being less cost and time-effective at the same time. Excellent for eukaryotic proteins, representing a natural cell habitat, protoplast isolation is widely used in protein interaction visualization techniques, like BiFC (Bimolecular fluorescence complementation) and FRET (Förster resonance energy transfer). In this protocol we present a another use of Arabidopsis protoplast in protein degradation assay, proving its high versatility as a tool in proteomics.
[摘要] 植物转化和外源蛋白质表达是分子生物学和生物技术的必要条件。 目前稳定植物转化的方法可能有问题且非常耗时。 因此，在原生质体中的瞬时表达已经成为有价值的替代方案，同时成本更低和时间有效。 优异的真核蛋白，代表天然细胞栖息地，原生质体分离广泛应用于蛋白质相互作用可视化技术，如BiFC（双分子荧光互补）和FRET（弗尔斯特共振能量转移）。 在本协议中，我们提出了在蛋白质降解测定中使用拟南芥原生质体，证明其作为蛋白质组学工具的高度通用性。
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