Use of native gels to measure protein binding to SSB

Jin Inoue; Tsutomu Mikawa

doi:10.1007/978-1-62703-032-8_13

Use of native gels to measure protein binding to SSB

Methods Mol Biol. 2012:922:175-82. doi: 10.1007/978-1-62703-032-8_13.

Authors

Jin Inoue¹, Tsutomu Mikawa

Affiliation

¹ Cellular & Molecular Biology Unit, RIKEN Advanced Science Institute, Yokohama, Japan.

PMID: 22976186
DOI: 10.1007/978-1-62703-032-8_13

Abstract

We describe a procedure to detect protein binding to SSB by polyacrylamide gel electrophoresis under non-denaturing conditions. As an example, we show the interaction of Thermus thermophilus (Tth) SSB with its cognate RecO protein. The interaction is detected as decay of the band corresponding to SSB by addition of RecO. We also demonstrate analysis of the RecO-RecR interaction as another example of this method.

MeSH terms

Bacterial Proteins / genetics
Bacterial Proteins / metabolism*
DNA-Binding Proteins / genetics
DNA-Binding Proteins / metabolism*
Electrophoresis, Polyacrylamide Gel / methods*
Molecular Biology / methods
Protein Binding
Recombination, Genetic
Thermus thermophilus / genetics
Thermus thermophilus / metabolism*

Substances

Bacterial Proteins
DNA-Binding Proteins