A freeze-thaw method for disintegration of Escherichia coli cells producing T7 lysozyme used in pBAD expression systems.

Comparison of T7- and Lac-Based Systems for the Periplasmic Expression of Human Granulocyte Macrophage Colony Stimulating Factor in Escherichia coli

CONSTRUCTION OF RECOMBINANT PLASMIDS FOR PERIPLASMIC EXPRESSION OF HUMAN GROWTH HORMONE IN ESCHERICHIA COLI UNDER T7 AND LAC PROMOTERS

In order to study the periplasmic expression of human growth hormone (hGH) in Escherichia coli, the related cDNA was inserted in two expression plasmids carrying pelB signal peptide, one with lac bacterial promoter and the other with a bacteriophage T7-based promoter. The recombinant plasmids were moved to TG1 and BL21 strains of E. coli, respectively. To induce the expression systems, IPTG and...

Cytoplasmic RNA Polymerase in Escherichia coli.

To obtain an estimate for the concentration of free functional RNA polymerase in the bacterial cytoplasm, the content of RNA polymerase beta and beta' subunits in DNA-free minicells from the minicell-producing Escherichia coli strain chi925 was determined. In bacteria grown in Luria-Bertani medium at 2.5 doublings/h, 1.0% of the total protein was RNA polymerase. The concentration of cytoplasmic...

Synthesis and Expression of Modified bFGF Gene in Escherichia coli Cells

A new strategy for construction of synthetic gene encoding human basic fibroblast growth factor comprising DNA annealing-ligation and augmentation by polymerase chain reaction was introduced. The sequence of the gene and corresponding amino acid chain were modified in order to increase stability of the protein. First, 300 bp and 160 bp fragments of the gene were assembled from 18 oligonucleotid...

The Inhibitory Effect of EDTA and Mg on the Activity of NADH Dehydrogenase in Lysozyme Lysis

NADH dehydrogenase of Escherichia coli, a component of the electron transport chain, oxidizes NADH to NAD. Previously, it was demonstrated that when E. coli K12 cells were lysed by the lysozyme lysis method, NADH dehydrogenase activity was inhibited; however, when cells were lysed by French press, NADH dehydrogenase activity was observed. In this study, components commonly used in lysozyme lysi...