Bacteriophage T7 late promoters: construction and in vitro transcription properties of deletion mutants.

AUTOR(ES)

Chapman, K A

RESUMO

The construction of plasmids containing T7 class I promoters with deletion mutants was described. Restriction fragments, ending at the Hinf I site located at position -10 in the promoter from 14.8% of the T7 genome, were cloned into pBR322. This produced the deletion of either the left or the right part of the promoter. The in vitro transcription properties of these plasmids were determined. Control plasmids were obtained by cloning wild type class II and class III promoters into pBR322. These plasmids also were used to compare the in vitro transcription properties of the two classes of late promoters. Much of the leftward part of a T7 late promoter can be deleted without abolishing activity, but deletion of the right part eliminates promoter activity. Class II, class III, and the mutated promoters have characteristic responses to changes in ionic strength, exogenous glycerol, and temperature.

Documentos Relacionados

• Construction of bacteriophage T7 late promoters with point mutations and characterization by in vitro transcription properties.
• M13 vectors with T7 polymerase promoters: transcription limited by oligonucleotides.
• Bacteriophage T7 late promoters with point mutations: quantitative footprinting and in vivo expression.
• Revised transcription map of the late region of bacteriophage T7 DNA.
• Detection of polycistronic and overlapping bacteriophage T7 late transcripts by in vitro translation.