Strategy for epitope tagging the protein-coding region of any gene.
We describe the construction of a plasmid carrying a DNA cassette encoding the influenza virus hemagglutinin (HA1) epitope. Insertion of this cassette into the coding region creates a gene encoding an epitope-tagged protein. The tagged protein can be detected and quantified by using a commercially available anti-HA1 antibody. A one-step procedure introduces the epitope at any restriction site within a gene while maintaining the open reading frame. An example of the application of this technique is presented and the different uses of the cassette are discussed.
Surdej
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P
Case Western Reserve University, School of Medicine, Cleveland, OH.
Jacobs-Lorena
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M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004331:Drosophila melanogaster; D000939:Epitopes / Q000235:genetics*; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral / Q000032:analysis / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D010957:Plasmids
D000939:Epitopes; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006389:Hemagglutinins, Viral