Specific labelling of the active site of T7 RNA polymerase.

AUTOR(ES)

Schäffner, A R

RESUMO

We describe a method for specifically labelling T7 RNA polymerase at (or near) the active site. Enzyme molecules that have been modified by covalent attachment of a benzaldehyde nucleotide derivative in the presence of template DNA are subsequently incubated with radioactively labelled nucleoside triphosphates. Labelling of the enzyme occurs as a result of the formation of the first phosphodiester bond. The labelling is template-directed and the expected specificity of initiation at individual T7 promoters is observed. The label has been localized to an 80 kd tryptic fragment that contains the carboxy-terminal portion of the enzyme.

Documentos Relacionados

• Transcription of RNA templates by T7 RNA polymerase.
• Preparation of capped RNA transcripts using T7 RNA polymerase.
• Rapid and simple purification of T7 RNA polymerase.
• Structure of a promoter for T7 RNA polymerase.
• RNA template-directed RNA synthesis by T7 RNA polymerase.