RNA-binding site in T7 RNA polymerase

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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...

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Nucleotide sequence of an RNA polymerase binding site at an early T7 promoter.

Escherichia coli RNA polymerase (EC 2.7.7.6), bound in a tight complex at an early T7 promoter, protects 41 to 43 base pairs of DNA from digestion by DNase. I. The protected DNA fragment contains both the binding site for RNA polymerase and the mRNA initiation point for the promoter. The sequence of the DNA fragment and the sequence of the mRNA that it codes for are presented here. A seven-base...

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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...

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Evolution of an inhibitory RNA aptamer against T7 RNA polymerase

Aptamers are promising gene components that can be used for the construction of synthetic gene circuits. In this study, we isolated an RNA aptamer that specifically inhibits transcription of T7 RNA polymerase (RNAP). The 38-nucleotide aptamer, which was a shortened variant of an initial SELEX isolate, showed moderate inhibitory activity. By stepwise doped-SELEX, we isolated evolved variants wit...

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ژورنال

عنوان ژورنال: Proceedings of the National Academy of Sciences

سال: 1998

ISSN: 0027-8424,1091-6490

DOI: 10.1073/pnas.95.16.9111