Identification of novel DNA-binding proteins using DNA-affinity chromatography/pull down.
Identification of novel DNA-binding proteins using DNA-affinity chromatography/pull down.
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DOI：
10.1002/9780471729259.mc01f01s24
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发表时间：
2012-02
影响因子：
--
通讯作者：
Stevenson, Brian
中科院分区：
文献类型：
--
作者：
Jutras, Brandon L;Verma, Ashutosh;Stevenson, Brian
提出了可以分离和鉴定新型细菌DNA结合蛋白的方法。简而言之，将感兴趣的DNA序列固定到珠上，然后与细菌细胞质提取物一起孵育。用含有非特异性DNA和低盐浓度的缓冲液洗涤将去除非粘附性和低特异性DNA结合蛋白，而随后用较高盐浓度的洗涤将去除更特异性的DNA结合蛋白。然后可以通过标准蛋白质组学技术鉴定洗脱的蛋白质。
Methods are presented through which one may isolate and identify novel bacterial DNA-binding proteins. Briefly, the DNA sequence of interest is affixed to beads, then incubated with bacterial cytoplasmic extract. Washes with buffers containing non-specific DNA and low salt concentrations will remove non-adhering and low-specificity DNA-binding proteins, while subsequent washes with higher salt concentrations will elute more specific DNA-binding proteins. Eluted proteins may then be identified by standard proteomic techniques.