Gene fusion vectors based on the gene for staphylococcal protein A |
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Authors: | Mathias Uhlén Björn Nilsson Bengt Guss Martin Lindberg Sten Gatenbeck Lennart Philipson |
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Affiliation: | 1. Department of Biochemistry, Royal Institute of Technology, S-100 44 StockholmSweden Tel. 8-7877513;2. Department of Microbiology, University of Uppsala, The Biomédical Center, Box 581, S-751 23 UppsalaSweden Tel. 18-174584 |
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Abstract: | Two plasmid vectors, containing the gene coding for staphylococcal protein A and adapted for gene fusion, have been constructed. These vectors will allow fusion of any gene to the protein A gene, thus giving hybrid proteins which can be purified, in a one-step procedure, by IgG affinity chromatography. As an example of the practical use of such vectors, the protein A gene has been fused to the lacZ gene of Escherichia coli. E. coli strains containing such plasmids produce hybrid proteins with both IgG binding and β-galactosidase activities. The hybrid protein(s) can be immobilized on IgG-Sepharose by its protein A moiety with high efficiency without losing its enzymatic activity and they can be eluted from the column by competitive elution with pure protein A. The fused protein(s) also binds to IgG-coated microtiter wells which means that the in vivo product can be used as an enzyme conjugate in ELISA tests. |
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Keywords: | Recombinant DNA hybrid protein IgG-binding β-galactosidase plasmids) bp base pairs ELISA see MATERIALS AND METHODS section c Fc constant part of immunoglobulins IgG immunoglobulin type G kb kilobase pairs ONPG PBST phosphate-buffered saline with 0 05% Tween 20 SDS sodium dodecyl sulfate Xgal |
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