Identification of novel DNA-binding proteins using DNA-affinity chromatography/pull down

使用 DNA 亲和层析/下拉法鉴定新型 DNA 结合蛋白

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作者:Brandon L Jutras, Ashutosh Verma, Brian Stevenson

Abstract

This units presents methods through which one may isolate and identify novel bacterial DNA-binding proteins. Briefly, the DNA sequence of interest is affixed to beads, and then incubated with bacterial cytoplasmic extract. Washes with buffers containing nonspecific 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.

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