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PMID: 3149606 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

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

Gene ·Vol. 73 ·No. 2 ·1988-12-20 ·Pages 305-18

Parmley SF, Smith GP

Abstract

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.

MeSH Terms
Antibodies Base Sequence Chromatography, Affinity Cloning, Molecular/methods Coliphages/drug effects,genetics DNA, Viral/genetics,immunology Escherichia coli/genetics Genes, Viral Genetic Vectors Molecular Sequence Data Transduction, Genetic Ultraviolet Rays Virion/genetics beta-Galactosidase/genetics
Chemicals
Antibodies DNA, Viral beta-Galactosidase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Parmley S F
Division of Biological Sciences, University of Missouri, Columbia 65211.
Smith G P
Article Info
Journal
Gene
Abbr.
Gene
ISSN
0378-1119
Published
1988-12-20
Pages
305-18
Language
English
Region
Netherlands
NLM ID
7706761
Subset
IM
Grants
NIAID NIH HHS · AI20564 · United States
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