Pyrosequencing of phage display libraries for the identification of cell-specific targeting ligands.
High combinatorial phage display libraries have become an important tool in the search for ligand-receptor interactions. The advantage this approach offers is the ability to screen large repertoires of peptides, displayed on the coat proteins of bacteriophages, against a target at the same time. In addition, no prior knowledge is required of the target or the ligand. However, to characterize the peptides of interest a short length of the bacteriophage genome that encodes the peptide sequence requires DNA sequencing. The number of candidate bacteriophages can be large and so sequencing is expensive, time-consuming, and laborious. Therefore, a methodology using Pyrosequencing has been developed where 96-phage displaying a seven amino acid peptide can be analyzed simultaneously within 45 min and at a fraction of the cost associated with traditional automated Sanger sequencing.
Rahim
Ahad A
AA
Gene Therapy Group, Chester Beatty Laboratories, Institute of Cancer Research, London, UK.
D001435:Bacteriophages / Q000235:genetics; D001483:Base Sequence; D004277:DNA, Single-Stranded / Q000235:genetics / Q000302:isolation & purification; D011756:Diphosphates / Q000378:metabolism*; D004926:Escherichia coli; D008024:Ligands; D008969:Molecular Sequence Data; D009928:Organ Specificity; D019151:Peptide Library; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods*; D013698:Templates, Genetic
D004277:DNA, Single-Stranded; D011756:Diphosphates; D008024:Ligands; D019151:Peptide Library