Site-directed mutagenesis in human granulocyte- colony stimulating factor, cloning and expression in Escherichia coli

Site-Directed Mutagenesis in Human Granulocyte-colony Stimulating Factor, Cloning and Expression in Escherichia coli

Human granulocyte colony stimulating factor (hG-CSF) induces proliferation and differentiation of granulocyte progenitor cells. This glycoprotein is currently being used for treatment of neutropenia, in patients who have undergone bone marrow transplantation. So far, different researchers have tried to enhance hG-CSF biological activity and stability. In this study, Polymerase Chain Reaction (P...

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

Expression and Secretion of Human Granulocyte Macrophage-Colony Stimulating Factor Using Escherichia coli Enterotoxin I Signal Sequence

With the aim of the secretion of human granulocyte macrophage-colony stimulating factor (hGM-CSF) in Escherichia coli, hGM-CSF cDNA was fused in-frame next to the signal sequence of ST toxin (ST-I) of exteroxigenic E. coli, containing 53 or 19 amino acids of signal peptide. The fused STsig::hGM-CSF coding fragments were inserted into a T7-based expression plasmid. The recombinant plasmids were ...

The Expression of Human Granulocyte Macrophage Colony Stimulating Factor by Heat-Induction in Escherichia coli

A self-regulated high-copy number plasmid containing chloramphenicol resistant gene, for the production of recombinant proteins under the regulation of bacteriophage ?pL promoter, was constructed. The designed 5024 base pair expression plasmid contained a heat sensitive repressor cI857 coding gene to regulate the function of ?pL promoter under heat shock induction. Using the constructed vector,...

Expression of a Chimeric Protein Containing the Catalytic Domain of Shiga-Like Toxin and Human Granulocyte Macrophage Colony-Stimulating Factor (hGM-CSF) in Escherichia coli and Its Recognition by Reciprocal Antibodies

Fusion of two genes at DNA level produces a single protein, known as a chimeric protein. Immunotoxins are chimeric proteins composed of specific cell targeting and cell killing moieties. Bacterial or plant toxins are commonly used as the killing moieties of the chimeric immunotoxins. In this investigation, the catalytic domain of Shiga-like toxin (A1) was fused to human granulocyte macrophage ...