Work

The detection of DNA-binding proteins by protein blotting

Year: 1980

Type: article

Abstract: A method, called "protein blotting," for the detection of DNA-binding proteins is described. Proteins are separated on an SDA-polyacrylamide gel. The gel is sandwiched between 2 nitrocellulose filters and the proteins allowed to diffuse out of the gel and onto the filters. The proteins are tightly bound to each filter, producing a replica of the original gel pattern. The replica is used to detect DNA-binding proteins, RNA-binding proteins or histone-binding proteins by incubation of the filter with [32P]DNA, [125I]RNA, or [125I] histone. Evidence is also presented that specific protein-DNA interactions may be detected by this technique; under appropriate conditions, the lac repressor binds only to DNA containing the lac operator. Strategies for the detection of specific protein-DNA interactions are discussed.

Source: Nucleic Acids Research

DOI: 10.1093/nar/8.1.1

Authors: Brian Bowen, Jay Steinberg, U.K. Laemmli, Harold Weintraub

Institutions: Princeton University, Fred Hutch Cancer Center

Language: English

Topic: Bacterial Genetics and Biotechnology

Subfield: Genetics

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: bronze

Funders: National Science Foundation, National Institutes of Health

Award: 1978 National Needs Postdoctoral Fellowship Program