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Genotype Phenotype Coupling pp 43–56Cite as

cDNA Display: A Stable and Simple Genotype–Phenotype Coupling Using a Cell-Free Translation System

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2070))

Abstract

A cDNA display method was developed based on the mRNA display method to increase its stability and efficiency for the directed evolution of various kinds of peptides and proteins. In this method, the puromycin-linker is a key molecule to realize smart genotype–phenotype coupling. A recently improved puromycin-linker and its use were explained in detail for the in vitro selection of peptides and proteins using the cDNA display method.

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References

  1. Nemoto N, Miyamoto-Sato E, Husimi Y, Yanagawa H (1997) In vitro virus: bonding of mRNA bearing puromycin at the 3′-terminal end to the C-terminal end of its encoded protein on the ribosome in vitro. FEBS Lett 414:405–408

    Article  CAS  Google Scholar 

  2. Roberts RW, Szostak JW (1997) RNA-peptide fusions for the in vitro selection of peptides and proteins. Proc Natl Acad Sci U S A 94:12297–12302

    Article  CAS  Google Scholar 

  3. Nishigaki K, Taguchi K, Kinoshita Y, Aita T, Husimi Y (1998) Y-ligation: an efficient method for ligating single-stranded DNAs and RNAs with T4 RNA ligase. Mol Divers 4:187–190

    Article  CAS  Google Scholar 

  4. Yamaguchi J, Naimuddin M, Biyani M, Sasaki T, Machida M, Kubo T, Funatsu T, Husimi Y, Nemoto N (2009) cDNA display: a novel screening method for functional disulfide-rich peptides by solid-phase synthesis and stabilization of mRNA-protein fusions. Nucleic Acids Res 37:e108

    Article  Google Scholar 

  5. Mochizuki Y, Biyani M, Tsuji-Ueno S, Suzuki M, Nishigaki K, Husimi Y, Nemoto N (2011) One-pot preparation of mRNA/cDNA display by a novel and versatile puromycin-linker DNA. ACS Comb Sci 13:478–485

    Article  CAS  Google Scholar 

  6. Mochizuki Y, Suzuki T, Fujimoto K, Nemoto N (2015) A versatile puromycin-linker using cnvK for high-throughput in vitro selection by cDNA display. J Biotechnol 212:174–180

    Article  CAS  Google Scholar 

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Author information

Authors and Affiliations

  1. Epsilon Molecular Engineering, Inc., Saitama, Japan

    Hidenao Arai, Shigefumi Kumachi & Naoto Nemoto

  2. Graduate School of Science and Engineering, Saitama University, Saitama, Japan

    Naoto Nemoto

Authors
  1. Hidenao Arai
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  2. Shigefumi Kumachi
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  3. Naoto Nemoto
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Corresponding author

Correspondence to Naoto Nemoto .

Editor information

Editors and Affiliations

  1. Protein Engineering and Antibody Technologies, Merck KGaA, Darmstadt, Germany

    Stefan Zielonka

  2. Protein Engineering and Antibody Technologies, Merck KGaA, Darmstadt, Germany

    Simon Krah

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Arai, H., Kumachi, S., Nemoto, N. (2020). cDNA Display: A Stable and Simple Genotype–Phenotype Coupling Using a Cell-Free Translation System. In: Zielonka, S., Krah, S. (eds) Genotype Phenotype Coupling. Methods in Molecular Biology, vol 2070. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9853-1_3

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  • DOI: https://doi.org/10.1007/978-1-4939-9853-1_3

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9852-4

  • Online ISBN: 978-1-4939-9853-1

  • eBook Packages: Springer Protocols

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