cDNA display technology
- What is cDNA display
- cDNA display is our protein display method which is genotype/phenotype linking systems. cDNA display has high stability and it handles high diversity.
- The Advantages of cDNA display
- Restriction of Displayed protein
- No restriction for proteins with cytotoxicity
- cDNA display enables to display proteins which have cytotoxicity for cells and microbes.
- Stability
- Very high stability
- Displayed protein is linking to cDNA display by linker. Therefore, cDNA display is very stable in alkaline pH, high temperature, and RNase.
- Diversity
- Very huge diversity
- Available to hundle approximately 10¹³⁻¹⁴
- Comparison of cDNA display and other protein display method
Display method Structure Genotype Diversity (/mL) Yeast display Plasmid DNA 10⁸ Phage display ssDNA 10⁸ cDNA display cDNA 10¹³⁻¹⁴ Ribosome display mRNA 10¹³ mRNA display mRNA 10¹³
- EME’s linker technology
- The method of genotype-phenotype linking strategy with puromycin linker
- After the preparation of the mRNA-cnvK linker, the mRNA-cnvK linker is added to cell free translation reaction mixture. Ribosome starts translation from 5’ ends side of mRNA, and polypeptides are synthesized. Because ribosome doesn’t translate the sequence of cnvK linker, ribosome stops translation when ribosome reaches the sequence of cnvK linker. At the same time, puromycin (aminoacyl tRNA analog) in the cnvK linker is integrated into ribosome. When the cnvK linker is integrated into ribosome, the peptide transfer reaction is occurred. Because of the peptide transfer reaction, the C-termini of synthesized polypeptide chain and puromycin linker is connected with covalent bond. Therefore, this synthesized compound is mRNA-cnvK linker polypeptide. In EME, we use our unique linker “photo-cross linkage puromycin linker (cnvK linker)” and “cDNA display synthesizer.” These technologies allow the auto preparation of large diversity (10¹³⁻¹⁴) of cDNA library.