High-level overproduction of His-tagged Tth DNA polymerase in Thermus thermophilus

Article Abstract:

A new plasmid is described, which uses a modified version of the promoter from the respiratory nitrate reductase (narp) to overproduce active DNA polymerase from Thermus thermophilus in larger amounts than those obtained with T7-dependent expression systems of Escherichia coli. It is concluded that this new expression system represents a competitive route for the direct overexpression of soluble enzymes in T. thermophilus, even compared to the high-level overexpression system of E. coli.

Plasmids

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Titration and conditional knockdown of the prfB gene in Escherichia ccoli: Effects on growth and overproduction of the recombinant mammalian selenoprotein thioredoxin reductase

Article Abstract:

An Escherichia coli strain is constructed that has endogenous chromosomal promoter of prfB replaced with the titratable [P.sub.BAD] promoter. The analysis has shown that expression of prfB can be titrated through targeted exchange of the native promoter with a [P.sub.BAD]-promoter and that knockdown of release factor 2 (RF2) has resulted in almost full efficiency of selenocysteine (Sec) incorporation at the cost of lower total seleno-protein yield.

Usage, Volumetric analysis, Chemical properties, Thioredoxin, Report

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Use of a dominant rpsL allele conferring streptomycin dependence for positive and negative selection in thermos thermophilus

Article Abstract:

A spontaneous rpsL mutant of Thermus thermophilus is isolated to search a new selection marker for this organism. Models built on the available three-dimensional structures of the 30S ribosomal subunit revealed that the K47R mutation directly affects the streptomycin binding site on S12.

Pharmaceutical Preparation Manufacturing, Pharmaceutical preparations, Streptomycin Preparations, Physiological aspects, Gene mutations, Gene mutation, Streptomycin

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