UV‐induced cross‐linking of proteins to plasmid pBR322 containing 8‐azidoadenine 2′‐deoxyribonucleotides

FEBS Letters

Dose, Klaus¹  Meffert, Rainer¹ 

[1] Institut für Biochemie, Johannes Gutenberg- Universität, Becherweg 30, D-6500 Mainz, FRG

关键词: Azidoadenine;    Plasmid pBR322;    DNA polymerase I;    Nick-translation;    DNA-protein cross-link;    Ultraviolet irradiation;    8-N3dATP;    8-azido-2′-deoxyadenosine-5′-triphosphate;    UV;    ultraviolet;   

DOI  :  10.1016/0014-5793(88)80914-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

An efficient method of cross-linking DNA to protein is described. The method is based on the incorporation of photoactive 8-azidoadenine 2′-deoxyribonucleotides into DNA. We have found that 8-N₃dATP is a substrate for E. coli DNA polymerase I and that 8-N₃dATP can be incorporated into plasmid pBR322 by nick-translation. Subsequently we were able to cross-link a set of different proteins to 8-azido-2′-deoxyadenosine-containing pBR322 by UV irradiation (366 nm). No DNA-protein photocross-linking was observed under the same conditions when the non-photoactive plasmid pBR322 was used.

【 授权许可】

Unknown