Trinucleotide cassettes increase diversity of T7 phage-displayed peptide library

Krumpe, Lauren RH; Schumacher, Kathryn M.; McMahon, James B.; Makowski, Lee; Mori, Toshiyuki

doi:10.1186/1472-6750-7-65

Trinucleotide cassettes increase diversity of T7 phage-displayed peptide library

Journal Article · Fri Oct 05 00:00:00 EDT 2007 · BMC Biotechnology (Online)

DOI:https://doi.org/10.1186/1472-6750-7-65· OSTI ID:1626539

Krumpe, Lauren RH ^[1]; Schumacher, Kathryn M. ^[2]; McMahon, James B. ^[3]; Makowski, Lee ^[4]; Mori, Toshiyuki ^[3]

National Cancer Institute, Frederick, MD (United States). Center for Cancer Research. Molecular Targets Development Program; National Cancer Institute, Frederick, MD (United States). SAIC-Frederick, Inc.; DOE/OSTI
National Cancer Institute, Frederick, MD (United States). Center for Cancer Research. Molecular Targets Development Program; National Cancer Institute, Frederick, MD (United States). Werner H. Kirsten Studen Internship Program
National Cancer Institute, Frederick, MD (United States). Center for Cancer Research. Molecular Targets Development Program
Argonne National Lab. (ANL), Argonne, IL (United States). Biosciences Division

Background: Amino acid sequence diversity is introduced into a phage-displayed peptide library by randomizing library oligonucleotide DNA. We recently evaluated the diversity of peptide libraries displayed on T7 lytic phage and M13 filamentous phage and showed that T7 phage can display a more diverse amino acid sequence repertoire due to differing processes of viral morphogenesis. Methods: In this study, we evaluated and compared the diversity of a 12-mer T7 phage-displayed peptide library randomized using codon-corrected trinucleotide cassettes with a T7 and an M13 12-mer phage-displayed peptide library constructed using the degenerate codon randomization method. Results: We herein demonstrate that the combination of trinucleotide cassette amino acid codon randomization and T7 phage display construction methods resulted in a significant enhancement to the functional diversity of a 12-mer peptide library. This novel library exhibited superior amino acid uniformity and order-of-magnitude increases in amino acid sequence diversity as compared to degenerate codon randomized peptide libraries. Comparative analyses of the biophysical characteristics of the 12-mer peptide libraries revealed the trinucleotide cassette-randomized library to be a unique resource. Conclusion: The combination of T7 phage display and trinucleotide cassette randomization resulted in a novel resource for the potential isolation of binding peptides for new and previously studied molecular targets.

View Accepted Manuscript (DOE)

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division; National Institutes of Health (NIH)

Grant/Contract Number:: AC02-06CH11357

OSTI ID:: 1626539

Journal Information:: BMC Biotechnology (Online), Journal Name: BMC Biotechnology (Online) Journal Issue: 1 Vol. 7; ISSN 1472-6750

Publisher:: BioMed CentralCopyright Statement

Country of Publication:: United States

Language:: English

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Compositional Bias in Naïve and Chemically-modified Phage-Displayed Libraries uncovered by Paired-end Deep Sequencing He, Bifang; Tjhung, Katrina F.; Bennett, Nicholas J. Scientific Reports, Vol. 8, Issue 1 https://doi.org/10.1038/s41598-018-19439-2	journal	January 2018
ProxiMAX randomization: a new technology for non-degenerate saturation mutagenesis of contiguous codons Ashraf, Mohammed; Frigotto, Laura; Smith, Matthew E. Biochemical Society Transactions, Vol. 41, Issue 5 https://doi.org/10.1042/bst20130123	journal	September 2013
TrimerDimer: an oligonucleotide-based saturation mutagenesis approach that removes redundant and stop codons Gaytán, Paul; Contreras-Zambrano, Casandra; Ortiz-Alvarado, Mónica Nucleic Acids Research, Vol. 37, Issue 18 https://doi.org/10.1093/nar/gkp602	journal	September 2009
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Construction of Nanobody Library in Mammalian Cells by Linear-double-stranded DNA Based AND Gate Genetic Circuit Zhang, Chunze; Tong, Xin; Li, Shuai BioRxiv https://doi.org/10.1101/2020.01.21.912907	posted_content	May 2020
DNA Libraries for the Construction of Phage Libraries: Statistical and Structural Requirements and Synthetic Methods Lindner, Thomas; Kolmar, Harald; Haberkorn, Uwe Molecules, Vol. 16, Issue 2 https://doi.org/10.3390/molecules16021625	journal	February 2011

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