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Catabolite-Insensitive Revertants of Lac Promoter Mutants

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TLDR
It is concluded that the promoter itself is the target site for both catabolite and transient repression of the lac operon.
Abstract
The maximum rate of expression of the lac operon is severely reduced in lac promoter mutants. Revertants of these mutations which produce higher levels of enzyme were isolated. Some of these revertants had lost sensitivity to catabolite repression and transient repression. The mutations responsible for these losses took place at sites very close to the original promoter mutations. From these results we conclude that the promoter itself is the target site for both catabolite and transient repression of the lac operon.

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Journal ArticleDOI

Recombinant protein expression in Escherichia coli: advances and challenges.

TL;DR: The different approaches for the synthesis of recombinant proteins in E. coli are reviewed and recent progress in this ever-growing field is discussed.
Journal ArticleDOI

A gel electrophoresis method for quantifying the binding of proteins to specific DNA regions: application to components of the Escherichia coli lactose operon regulatory system

TL;DR: It is demonstrated that even when pre-formed in the presence of CAP-cAMP, the polymerase-promoter open complex becomes unstable if CAP is then selectively removed, and this gel method is applied to the study of the E. coli lactose operon regulatory system.
PatentDOI

Cloning and expression of the gene for bacteriophage T7 RNA polymerase

TL;DR: In this paper, the authors describe a method to clone a functional gene for bacteriophage T7 RNA polymerase, which is useful for synthesizing large amounts of RNA in vivo or in vitro, and can produce a single RNA selectively from a complex mixture of DNAs.
Journal ArticleDOI

Tuning Escherichia coli for membrane protein overexpression

TL;DR: A derivative strain of E. coli BL21(DE3) is engineered, termed Lemo21(de3), in which the activity of the T7 RNA polymerase can be precisely controlled by its natural inhibitor T7 lysozyme (T7Lys), which allows optimizing overexpression of any given membrane protein by using only a single strain rather than a multitude of different strains.
Journal ArticleDOI

Coordination of bacterial proteome with metabolism by cyclic AMP signalling

TL;DR: A quantitative physiological approach is used to show that cAMP signalling tightly coordinates the expression of catabolic proteins with biosynthetic and ribosomal proteins, in accordance with the cellular metabolic needs during exponential growth, to ensure that proteomic resources are spent on distinct metabolic sectors as needed in different nutrient environments.
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