TY - JOUR
T1 - Identification of Novel DNA-Binding proteins Using DNA-Affinity Chromatography/Pull Down
AU - Jutras, Brandon L.
AU - Verma, Ashutosh
AU - Stevenson, Brian
PY - 2012/2
Y1 - 2012/2
N2 - 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 DNAbinding 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.
AB - 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 DNAbinding 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.
KW - DNA-binding protein
KW - Gene regulation
KW - Prokaryote
UR - https://www.scopus.com/pages/publications/84861306825
UR - https://www.scopus.com/pages/publications/84861306825#tab=citedBy
U2 - 10.1002/9780471729259.mc01f01s24
DO - 10.1002/9780471729259.mc01f01s24
M3 - Article
C2 - 22307548
AN - SCOPUS:84861306825
SN - 1934-8525
JO - Current Protocols in Microbiology
JF - Current Protocols in Microbiology
IS - SUPPL.24
M1 - 1F.1
ER -