Expression of an antibody-targeted plasminogen activator in Escherichia coli using chemically induced stress responses.
A recombinant gene coding for an antibody-targeted urokinase-type plasminogen activator was constructed for the purpose of enhancing the thrombolytic specificity of urokinase. The recombinant gene was cloned into prokaryotic expression vector pTrcHisA, and transformed into Escherichia coli strain Rosetta (DE3). Less than 4 mg of the desired protein l(-1) could be obtained in the form of inclusion bodies. Of various inducers and enhancers of stress responses, the heat-shock enhances, streptomycin, the osmotic stress inducers, D-arabinose and sucrose, and the cold-shock enhancer, tetracycline, simulated the expression of the antibody-targeted plasminogen activator by 2-5-fold.
Yu
Ming
M
National Key Laboratory of Protein Engineering, College of Life Sciences, Peking University, Beijing, 100871, P. R. China.
Yu
Meimin
M
Ru
Binggen
B
D000911:Antibodies, Monoclonal / Q000235:genetics / Q000378:metabolism; D002241:Carbohydrates / Q000494:pharmacology; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D000431:Ethanol / Q000494:pharmacology; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D005990:Glycerol / Q000494:pharmacology; D018869:Heat-Shock Response / Q000187:drug effects / Q000502:physiology*; D009997:Osmotic Pressure / Q000187:drug effects; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D013307:Streptomycin / Q000494:pharmacology; D013752:Tetracycline / Q000494:pharmacology; D014568:Urokinase-Type Plasminogen Activator / Q000096:biosynthesis* / Q000235:genetics*
D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D011993:Recombinant Fusion Proteins; D000431:Ethanol; D014568:Urokinase-Type Plasminogen Activator; D013752:Tetracycline; D005990:Glycerol; D013307:Streptomycin