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Integral water purification system

整体式水净化系统

Company: EMD Millipore
Catalog#: Milli-Q+
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An Assay to Test the Capacity of Arabidopsis Plant Defensin Type1 Protein to Induce Cellular Zinc (Zn) Tolerance in Yeast
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2015-11-20
[Abstract]  Heterologous expression of genes in budding yeast Saccharomyces cerevisiae (S. cerevisiae) is especially suitable to functionally study the corresponding encoded protein at the cellular level (Bonneaud et al., 1991). This is mainly because many strains defective in specific activities are available and could be complemented by homologous genes existing across the eukaryotic kingdom (http://www.yeastgenome.org/). However, the protocol we describe here is not a complementation but a “gain-of-function” assay. It is based on a drop-test assay that we have set up to assess the cellular zinc tolerance conferred by the expression of heterologous genes in the wild-type S. cerevisiae. Different dilutions ... [摘要]  基因在出芽酵母酿酒酵母(酿酒酵母)中的异源表达特别适合于在细胞水平上功能性研究相应的编码蛋白质(Bonneaud等人, ,1991)。这主要是因为许多具有特异性活性缺陷的菌株是可获得的,并且可以通过真核生物界存在的同源基因来补充( http: /www.yeastgenome.org/)。然而,我们在这里描述的协议不是互补,而是"获得功能"测定。它是基于滴试验测定,我们已经设置以评估由异源基因在野生型S中的表达赋予的锌细胞耐受性。酿酒厂。将表达目的异源基因的酵母培养物的不同稀释液在一系列富锌平板上生长,然后与表达空载体的对照酵母进行比较。使用不同浓度的酵母和锌对于在酵母转化后成功描述锌耐受性表型是必要的(Mirouze等人,2006)。该试验也已经证明对于区分基因家族的相关成员是有价值的,如拟南芥属植物防御素类型1所例证的(Shahzad等人,2013)。

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