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

Hidenao Arai; Shigefumi Kumachi; Naoto Nemoto

doi:10.1007/978-1-4939-9853-1_3

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

Methods Mol Biol. 2020:2070:43-56. doi: 10.1007/978-1-4939-9853-1_3.

Authors

Hidenao Arai¹, Shigefumi Kumachi¹, Naoto Nemoto^{2

3}

Affiliations

¹ Epsilon Molecular Engineering, Inc., Saitama, Japan.
² Epsilon Molecular Engineering, Inc., Saitama, Japan. nemoto@mail.saitama-u.ac.jp.
³ Graduate School of Science and Engineering, Saitama University, Saitama, Japan. nemoto@mail.saitama-u.ac.jp.

PMID: 31625089
DOI: 10.1007/978-1-4939-9853-1_3

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.

Keywords: Directed evolution; In vitro selection; Photocrosslinking; Puromycin-linker; cDNA display; cnvK; mRNA display.

MeSH terms

Animals
Cell-Free System / chemistry
Cell-Free System / metabolism
DNA, Complementary* / chemistry
DNA, Complementary* / genetics
DNA, Complementary* / metabolism
Protein Biosynthesis*
RNA, Messenger* / chemistry
RNA, Messenger* / genetics
RNA, Messenger* / metabolism
Rabbits

Substances

DNA, Complementary
RNA, Messenger