A Novel Approach to Peptide/Protein Chip Fabrication
This technology describes improved methods for stalling ribosomes during protein translation and capturing the nascent polypeptides through puromycin which is grafted at the end of an oligo immobilized on a solid phase. Existing technologies include an inefficient ligation step to incorporate puromycin onto the nucleic acid molecule, which is not required with this approach. This technology eliminates the need for the time consuming and labor intensive step of cloning ORFs into expression vectors as proteins can be generated directly from mRNA preparations or PCR templates in vitro. This makes protein chip production more amenable to high throughput production. Description (Set) Proposed Use (Set) Protein chips are emerging as a very powerful tool for proteomic studies. This technology provides new methods for synthesizing proteins chips at very high density from in vitro translated proteins. Furthermore, as the proteins do not require affinity tags for attachment, native proteins are immobilized uniformly onto the chips, making them more suitable for functional analyses.
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