Synthesis of mixed ribo/deoxyribopolynucleotides by mutant T7 RNA polymerase

Enzymatic RNA Synthesis using Bacteriophage T7 RNA Polymerase

Inhibition of T7 RNA polymerase by T7 lysozyme in vitro.

The in vivo observation that the expression of bacteriophage T7 gene 3.5 (T7 lysozyme) inactivates T7 class II transcription and the in vitro observation that T7 lysozyme inhibits T7 RNA polymerase lead to the hypothesis that T7 lysozyme might preferentially inhibit transcription from T7 class II promoters. T7 lysozyme was cloned into a lambda pL expression vector, overproduced in Escherichia c...

Transcription of RNA templates by T7 RNA polymerase.

Although highly specialized, T7 RNA polymerase seems to possess a large range of DNA- and RNA-dependent properties. To study such flexibility, we determined the ability of T7 RNA polymerase to transcribe chimeric DNA-RNA and RNA templates following initiation at a double stranded DNA promoter. We have found that T7 RNA polymerase is able to initiate on RNA templates, was processive, and was abl...

Kinetic mechanism of GTP binding and RNA synthesis during transcription initiation by bacteriophage T7 RNA polymerase.

We have used stopped-flow and rapid chemical quench-flow methods to investigate the kinetics of the early steps during transcription initiation by bacteriophage T7 RNA polymerase. Most promoters of T7 RNA polymerase initiate with two GTPs. The kinetics of GTP binding was investigated by monitoring the fluorescence changes resulting from GTP binding to polymerase and fluorescent 2-aminopurine-co...

RNA-binding site in T7 RNA polymerase.

Recent models of RNA polymerase transcription complexes have invoked the idea that enzyme-nascent RNA contacts contribute to the stability of the complexes. Although much progress on this topic has been made with the multisubunit Escherichia coli RNA polymerase, there is a paucity of information regarding the structure of single-subunit phage RNA polymerase transcription complexes. Here, we pho...