Cloning and expression of the gene for bacteriophage T7 RNA polymerase.

Bacteriophage T7 RNA polymerase-based expression in Pichia pastoris.

A novel Pichia pastoris expression vector (pEZT7) for the production of recombinant proteins employing prokaryotic bacteriophage T7 RNA polymerase (T7 RNAP) (EC 2.7.7.6) and the corresponding promoter pT7 was constructed. The gene for T7 RNAP was stably introduced into the P. pastoris chromosome 2 under control of the (endogenous) constitutive P. pastoris glyceraldehyde-3-phosphate dehydrogenas...

Enzymatic RNA Synthesis using Bacteriophage T7 RNA Polymerase

The structure of bacteriophage T7 lysozyme, a zinc amidase and an inhibitor of T7 RNA polymerase.

The lysozyme of bacteriophage T7 is a bifunctional protein that cuts amide bonds in the bacterial cell wall and binds to and inhibits transcription by T7 RNA polymerase. The structure of a mutant T7 lysozyme has been determined by x-ray crystallography and refined at 2.2-A resolution. The protein folds into an alpha/beta-sheet structure that has a prominent cleft. A zinc atom is located in the ...

Physical mapping of hybrid bacteriophage T7/T3 RNA polymerase genes.

The late regions of the T7 and T3 bacteriophage genomes are transcribed by phage-specified RNA polymerases, the products of gene 1. Although these phage transcriptional systems share many characteristics and are obviously related, they have diverged to such an extent that neither of their respective RNA polymerases utilizes the promotor sites of the other phage at an appreciable rate. However, ...

Transcription elongation past O6-methylguanine by human RNA polymerase II and bacteriophage T7 RNA polymerase

O(6)-Methylguanine (O(6)-meG) is a major mutagenic, carcinogenic and cytotoxic DNA adduct produced by various endogenous and exogenous methylating agents. We report the results of transcription past a site-specifically modified O(6)-meG DNA template by bacteriophage T7 RNA polymerase and human RNA polymerase II. These data show that O(6)-meG partially blocks T7 RNA polymerase and human RNA poly...