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 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.
Krebber
A
A
Biochemisches Institut der Universität Zürich, Switzerland.
Burmester
J
J
Plückthun
A
A
D001426:Bacterial Proteins / Q000235:genetics; D017104:Bacteriophage M13 / Q000235:genetics; D001483:Base Sequence; D036022:Capsid Proteins; D003001:Cloning, Molecular / Q000379:methods*; D004274:DNA, Recombinant; D004268:DNA-Binding Proteins / Q000235:genetics*; D029968:Escherichia coli Proteins; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D007128:Immunoglobulin Fragments / Q000235:genetics; D056968:Lac Repressors; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000494:pharmacology; D012097:Repressor Proteins / Q000235:genetics; D013728:Terminator Regions, Genetic; D014760:Viral Fusion Proteins / Q000235:genetics*
D001426:Bacterial Proteins; D036022:Capsid Proteins; D004274:DNA, Recombinant; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D007128:Immunoglobulin Fragments; D056968:Lac Repressors; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D014760:Viral Fusion Proteins; C055966:immunoglobulin Fv