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文献和实验了不同的内含肽2,pTWIN1用Mxe GyrA 内含肽(198 aa)作为内含肽2;而pTWIN2使用的是Mth RIR1 内含肽(134 aa)。然而,pTWIN1 和pTWIN2含有相同的多克隆位点,这样简化了将目的基因插入两个载体来选择最佳表达质粒的操作。pTWIN-MBP融合了麦芽糖蛋白,可用作对照载体和克隆载体(将目的基因克隆到pTWIN-MBP1 NcoI-SacI酶切位点之间,会在蛋白产物的N-端增加3个氨基酸,C-端增加23个氨基酸。)IMPACT系统利用的是T7启动子进行转录
Purification of Human Multiprotein Complexes using OneSTrEP Technology
. For more theoretical background and a protocol for purification of recombinant Strep -tag® fusion proteins from E.coli please refer to Schmidt and Skerra (2007). In comparison to other commonly used purification strategies (i.e. HA and FLAG double-tag
Purification of Human Multiprotein Complexes using OneSTrEP Technology (PROT41)
complexes from human HeLa S3 cells in a scale and purity optimized for characterization by mass spectrometry. For this purpose, we use stably expressed One-STrEP -tag ® fusion proteins. This approach was successfully used in characterization of histone
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