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Totally in vitro protein selection using mRNA-protein fusions and ribosome display

✍ Scribed by Richard W Roberts

Publisher
Elsevier Science
Year
1999
Tongue
English
Weight
751 KB
Volume
3
Category
Article
ISSN
1367-5931

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✦ Synopsis

Both chemists and biochemists have great interest in creating peptides and proteins with desired structure, recognition and catalytic properties. Recently, two techniques have been developed that afford functional selection of proteins entirely in vitro: mRNA-protein fusions, and ribosome display. Improvements in both methods have allowed peptide and protein libraries of unprecedented size (1011-l 01s different members) to be generated and screened for function.

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Protein–Ribosome–mRNA Display: Affinity Isolation of Enzyme–Ribosome–mRNA Complexes and cDNA Cloning in a Single-Tube Reaction
✍ Erhard Bieberich; Dmitri Kapitonov; Tewin Tencomnao; Robert K. Yu 📂 Article 📅 2000 🏛 Elsevier Science 🌐 English ⚖ 82 KB

An enzyme-ribosome-mRNA complex was specifically purified by binding to the immobilized enzyme substrate and the cDNA was cloned in a single-tube reaction by one-step reverse transcription-PCR. The ganglioside GM3, used by sialyltransferase II (ST-II) as a substrate, was coated on a 96-well microtit