CONSTRUCTION OF RECOMBINANT ESCHERICHIA-C OLI STRAINS FOR SECRETORY EXPRESSION OF ARTIFICIAL GENES FOR HUMAN GRANULOCYTE-MACROPHAGE COLONY-STIMULATING FACTOR

A number of recombinant plasmids for expression of artificial genes en coding human granulocyte-macrophage colony stimulating factor (GM-CSF) were constructed. A hybrid gene was obtained that contains a sequence encoding the leader peptide and a tandem of two IgG-binding domains o f protein A from Staphylococcus aureus coupled, through an enteropepdi dase linker, to a synthetic gmcsf gene. The construction enables Esche richia coil to carry out biosynthesis of the hybrid protein and its su bsequent transport into the periplasmic space of bacteria. Another hyb rid gene, combining sequences for the signal peptide of the E. coli ou ter membrane protein OmpA and GM-CSF, was obtained using polymerase ch ain reaction. The localization of the mature protein produced by the h ybrid gene was found to depend on the strength of the promoter used.