Detection of E. coli colonies expressing the v-sis oncogene product with monoclonal antibodies made against synthetic peptides.
A method for immunodetection of individual epitopes on eukaryotic proteins synthesized in E. coli colonies is described. The system is developed using monoclonal antibodies produced against the Ha-ras protein produced in E. coli JM103. Monoclonal antibodies made against a synthetic peptide from the v-sis oncogene sequence are then used to identify bacterial colonies in which the v-sis protein is being produced. Production of the v-sis protein by these E. coli colonies was confirmed by immunoblot analysis. The assay utilizes peroxidase conjugated anti-mouse immunoglobulin and 4-chloro-1-naphthol to detect the positive colonies and can detect on the order of 200 pg antigen per E. coli colony.
Kennett
R H
RH
Leunk
R
R
Meyer
B
B
Silenzio
V
V
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology; D000939:Epitopes; D004926:Escherichia coli / Q000235:genetics; D007124:Immunoenzyme Techniques; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000276:immunology*; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000939:Epitopes; D009856:Oncogene Proteins, Viral; D010446:Peptide Fragments; D011994:Recombinant Proteins