Patent Document ID: 10011830
Application ID: 14890864
Patent Flag: 1

Claim One:
1. A method for displaying a polypeptide on a DNA template encoding it, the method comprising: a) providing a DNA template that encodes the polypeptide, wherein the DNA template comprises: i) a promoter operably linked to an open reading frame (ORF); and ii) a molecular roadblock that blocks progress of an RNA polymerase during transcription of the DNA template, wherein the roadblock is located at the 5′ end of the DNA template antisense strand, or at the 3′ end of the DNA template sense strand, or at both the 5′ end of the DNA template antisense strand and the 3′ end of the DNA template sense strand; b) adding an RNA polymerase that can be blocked by the molecular roadblock, wherein the RNA polymerase binds to the promoter of the DNA template and carries out transcription of the DNA template until the RNA polymerase stalls at the molecular roadblock where the RNA polymerase is blocked from further progress, such that the DNA template and transcribed mRNA remain associated; and c) adding a ribosome that binds to a ribosomal binding site on the mRNA; d) carrying out translation under conditions of amino acid starvation, wherein the ribosome stalls at a particular position on the RNA transcript because the amino acid needed for addition at that position is not provided; e) removing any unbound ribosomes; and f) adding the missing amino acid needed to resume translation, which continues until the one bound ribosome reaches the stalled RNA polymerase at the molecular roadblock where the ribosome is blocked from further progress, such that the ribosome displays the nascent polypeptide while remaining associated with the mRNA.