A preparative method for crosslinking proteins to DNA in nuclei by single-pulse UV laser irradiation.
Photochem Photobiol
; 66(1): 42-5, 1997 Jul.
Article
em En
| MEDLINE
| ID: mdl-9230703
Photocrosslinking of proteins to DNA by single-pulse UV laser has been used only in analytical experiments, carried out with reconstituted complexes of a single DNA binding protein and a labeled target sequence. Here we propose a large-scale technique for irradiation of nuclei, generating preparative quantities of covalently linked protein-DNA complexes for further analysis of the partner molecules. The use of a flow cuvette allows a milligram of DNA in either nuclei or chromatin to be irradiated by a single pulse within few minutes. The efficiency of crosslinking varies from 6 to 12% of the total nuclear proteins. The presence of histones and other chromosomal proteins in the crosslinked protein-DNA complexes was demonstrated by using specific antibodies. The irradiation procedure can be fully automated using a microcomputer.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Raios Ultravioleta
/
DNA de Neoplasias
/
Histonas
Limite:
Animals
Idioma:
En
Revista:
Photochem Photobiol
Ano de publicação:
1997
Tipo de documento:
Article
País de afiliação:
Bulgária