MRNA display
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- The correct title of this article is mRNA display. The initial letter is shown capitalized due to technical restrictions.
mRNA display is a display technique used to perform in vitro protein, and/or peptide evolution to create molecules that can bind to a desired ligand. The process results in translated peptides or proteins that are associated with their mRNA progenitor, via a puromycin linkage, which is used, as a complex, to bind to an immobilized ligand in a selection step (affinity chromatography). The mRNA-protein fusions that bind well are then reverse transcribed to cDNA and their sequence amplified via a polymerase chain reaction. The end result is a nucleotide sequence that encodes peptides which tightly bind molecules of interest.
[edit] Technical details
A synthetic mRNA with puromycin at its 3' end forms an mRNA-peptide fusion that is purified by affinity chromatography. By linking the peptide to its own mRNA, a molecular rosetta stone is created.
Competing methods for protein evolution in vitro are phage display, yeast display, bacterial display, and ribosome display.
[edit] Citations
- Roberts, R.W., Szostak, J.W.; Proc. Natl. Acad. Sci. U.S.A., 1997, 94, 12297-12302.
- Keefe, A.D., Szostak, J.W.; Nature, 2001, 410, 715-18.
[edit] Notes
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