Production of the chimeric-binding protein, maltose-binding protein-protein A, by gene fusion.
A fusion protein between maltose-binding protein (MBP) and staphylococcal protein A (SpA) was genetically produced. The gene fusion plasmid, pMALPA2, was constructed by inserting the protein A gene into an expression vector of maltose-binding protein in frame, and was expressed efficiently in Escherichia coli. The resulting fusion protein of molecular mass 65 kDa, retained the activity of both MBP and SpA (binding capability to amylose and immunoglobulin G). This chimeric-binding protein was used as an adhesive molecule for immobilization of antibodies to a solid-phase surface for enzyme immunoassay. An enzyme immunoassay was performed with the fusion protein, and human IgG was determined in the concentration range from 10(-4) to 10(-6) g ml-1.
Kobatake
E
E
Department of Bioengineering, Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology Nagatsuta, Yokohama, Japan.
Ikariyama
Y
Y
Aizawa
M
M
D018528:ATP-Binding Cassette Transporters; D002352:Carrier Proteins / Q000235:genetics*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000032:analysis; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000235:genetics; D013205:Staphylococcal Protein A / Q000235:genetics*
D018528:ATP-Binding Cassette Transporters; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D007074:Immunoglobulin G; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; C461599:maltose transport system, E coli