Immunocytochemical demonstration of human proinsulin chimeric polypeptide within cytoplasmic inclusion bodies of Escherichia coli.
Immunocytochemical techniques were used to identify human proinsulin chimeric protein in cytoplasmic inclusion bodies of genetically modified Escherichia coli. Antibodies to proinsulin chimeric protein (human proinsulin coupled at its amino-terminus to a portion of the E. coli tryptophan E gene product) were localized in E. coli using post-embedding staining with protein A-peroxidase labelling for transmission electron microscopy. The observable distribution of the labelled antibody was limited to that portion of the E. coli cytoplasm occupied by inclusion bodies. The localization of human peptides as insoluble masses within the bacterial cytoplasm has important implications in relation to the synthesis, recovery and purification of pharmacologically useful substances produced through the application of recombinant DNA technology.
D001426:Bacterial Proteins / Q000032:analysis*; D004926:Escherichia coli / Q000378:metabolism / Q000648:ultrastructure*; D006651:Histocytochemistry; D006801:Humans; D007124:Immunoenzyme Techniques; D002479:Inclusion Bodies / Q000032:analysis*; D008854:Microscopy, Electron; D011384:Proinsulin / Q000032:analysis*; D011993:Recombinant Fusion Proteins
D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; C039739:proinsulin-E. coli tryptophan E chimeric polypeptide; D011384:Proinsulin