Ward A C, Castelli L A, Macreadie I G, Azad A A
Biomolecular Research Institute, Parkville, Victoria, Australia.
Yeast. 1994 Apr;10(4):441-9. doi: 10.1002/yea.320100403.
We describe six new yeast episomal vectors which encode glutathione S-transferase (GST) affinity tags. These allow for the production of GST-fusion proteins in Saccharomyces cerevisiae under the control of the CUP1 promoter. Affinity chromatography with glutathione-Sepharose permits convenient purification of the fusion protein from a yeast lysate. The presence of a protease cleavage site facilitates subsequent removal of the GST tag. The expression and single-step purification of both GST and a functional GST-metallothionein fusion from yeast are shown as an example of the application of these vectors.
我们描述了六种编码谷胱甘肽S-转移酶(GST)亲和标签的新型酵母附加型载体。这些载体可在CUP1启动子的控制下,于酿酒酵母中生产GST融合蛋白。用谷胱甘肽-琼脂糖进行亲和层析可方便地从酵母裂解物中纯化融合蛋白。蛋白酶切割位点的存在便于随后去除GST标签。以从酵母中表达并单步纯化GST以及一种功能性GST-金属硫蛋白融合蛋白为例,展示了这些载体的应用。