Identification and characterization of mutant clones with enhanced propagation rates from phage-displayed peptide libraries.
Anal Biochem
; 462: 35-43, 2014 Oct 01.
Article
en En
| MEDLINE
| ID: mdl-24952360
ABSTRACT
A target-unrelated peptide (TUP) can arise in phage display selection experiments as a result of a propagation advantage exhibited by the phage clone displaying the peptide. We previously characterized HAIYPRH, from the M13-based Ph.D.-7 phage display library, as a propagation-related TUP resulting from a GâA mutation in the Shine-Dalgarno sequence of gene II. This mutant was shown to propagate in Escherichia coli at a dramatically faster rate than phage bearing the wild-type Shine-Dalgarno sequence. We now report 27 additional fast-propagating clones displaying 24 different peptides and carrying 14 unique mutations. Most of these mutations are found either in or upstream of the gene II Shine-Dalgarno sequence, but still within the mRNA transcript of gene II. All 27 clones propagate at significantly higher rates than normal library phage, most within experimental error of wild-type M13 propagation, suggesting that mutations arise to compensate for the reduced virulence caused by the insertion of a lacZα cassette proximal to the replication origin of the phage used to construct the library. We also describe an efficient and convenient assay to diagnose propagation-related TUPS among peptide sequences selected by phage display.
Palabras clave
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Péptidos
/
Biblioteca de Péptidos
/
Mutación
Tipo de estudio:
Diagnostic_studies
Idioma:
En
Revista:
Anal Biochem
Año:
2014
Tipo del documento:
Article
País de afiliación:
Estados Unidos