Antibody-selectable filamentous fd phage vectors: affinity purification of target genes

Foreign DNA fragments can be inserted into a minor coat protein gene of filamentous phage, creating a fusion protein that is incorporated into the virion; we call these particles “fusion phage”. The foreign amino acids are displayed on the surface, allowing fusion phage bearing antigenic determinant...

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Veröffentlicht in:Gene 1988-12, Vol.73 (2), p.305-318
Hauptverfasser: Parmley, Stephen F., Smith, George P.
Format: Artikel
Sprache:eng
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Zusammenfassung:Foreign DNA fragments can be inserted into a minor coat protein gene of filamentous phage, creating a fusion protein that is incorporated into the virion; we call these particles “fusion phage”. The foreign amino acids are displayed on the surface, allowing fusion phage bearing antigenic determinants from a target gene to be purified in infectious form by affinity to antibody directed against the gene product. Here we introduce fusion-phage vectors that accept foreign DNA inserts with little effect on phage function; and describe affinity purification of virions bearing a target determinant from a 10 8-fold excess of phage not bearing the determinant, using minute amounts of antibody. These “antibody-selectable” vectors are a promising alternative to conventional expression systems for using antibodies to clone genes, though the ability to isolate rare clones from actual libraries remains to be demonstrated.
ISSN:0378-1119
1879-0038
DOI:10.1016/0378-1119(88)90495-7