Inclusion of an upstream transcriptional terminator in phage display vectors abolishes background expression of toxic fusions with coat protein g3p

Expression of toxic gene products affects bacterial cell growth and phage display, causing a strong selection against plasmid maintenance and integrity. During phage propagation steps, in particular, phagemid instability can dramatically affect diversity of antibody libraries or even lead to the del...

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Bibliographic Details
Published in:Gene 1996-10, Vol.178 (1), p.71-74
Main Authors: Krebber, Anke, Burmester, Jörg, Plückthun, Andreas
Format: Article
Language:English
Subjects:
Antibody libraries
Bacterial Proteins - genetics
Bacteriophage M13 - genetics
Base Sequence
Capsid Proteins
Cloning, Molecular - methods
DNA, Recombinant
DNA-Binding Proteins - genetics
E. coli
Escherichia coli Proteins
Filamentous phage
Gene Expression Regulation
Genetic Vectors
Immunoglobulin Fragments - genetics
Lac Repressors
Molecular Sequence Data
Promoter Regions, Genetic
Recombinant antibodies
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - pharmacology
Repressor Proteins - genetics
Terminator Regions, Genetic
Viral Fusion Proteins - genetics
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Summary:Expression of toxic gene products affects bacterial cell growth and phage display, causing a strong selection against plasmid maintenance and integrity. During phage propagation steps, in particular, phagemid instability can dramatically affect diversity of antibody libraries or even lead to the deletion of antibody genes. We constructed a modified phage display vector by introducing a strong transcriptional terminator upstream of the lac promoter, which, together with glucose suppression of its CAP-dependent activation, very efficiently represses product formation before induction.
ISSN:0378-1119
1879-0038
DOI:10.1016/0378-1119(96)00337-X