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Ribosome complexes as selection particles for in vitro development and evolution of proteins.
Ribosome complexes as selection particles for in vitro development and evolution of proteins.
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机译:核糖体复合物作为蛋白质体外发育和进化的选择颗粒。
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摘要
The invention provides a method to display peptides or nascent proteins as complex with ribosomes eukaryotic and mRNA encoding the protein or peptide following transcription and translation in vitro, to select complex bearing a peptide or nascent protein by of binding to a ligand, antigen or antibody, and subsequently recovering the genetic information encoding the peptide or protein complex selected ribosomes by reverse reaction and the polymerase chain reaction (RT-PCR) transcription. The recovery step RT-PCR is carried out directly on the intact ribosome complex, without prior dissociation to release the mRNA, thus contributing to their high efficiency and sensitivity. The steps of display, selection and recovery can be repeated in consecutive cycles. The method is exemplified using constructs such as single chain antibodies antibody-ribosome complex-mRNA (ARM).
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