Using a nucleic acid construct, association of a polypeptide with a sequence coding therefor and screening of a polypeptide that binds to a target substance are carried out, which nucleic acid construct comprises a 5-untranslated region and a coding region, wherein the above-mentioned coding region comprises a sequence coding for a polypeptide subjected to be displayed, a sequence coding for a first nucleic acid binding polypeptide, and a sequence coding for a second nucleic acid binding polypeptide; the above-mentioned 5-untranslated region comprises a first sequence capable of binding to a first nucleic acid binding polypeptide and a second sequence capable of binding to second nucleic acid binding polypeptide; and, when the above-mentioned nucleic acid construct is introduced in a translation system, a fusion protein translated from the coding region of the above-mentioned nucleic acid construct forms a complex with an RNA corresponding to the above-mentioned nucleic acid construct.

The polynucleotide construct of (1) or (2) below is used to perform ribosome display, CIS display and/or mRNA display in order to screen a Fab against an antigen of interest: (1) a polynucleotide construct which monocistronically comprises a ribosome-binding sequence, Fab first chain-coding sequence, linker peptide-coding sequence, Fab second chain-coding sequence and scaffold-coding sequence in this order, and further comprises at its 3-end a structure necessary for maintaining a complex with the Fab encoded by itself; and (2) a polynucleotide construct which comprises a Fab first chain-expressing cistron and a Fab second chain-expressing cistron each containing a ribosome-binding sequence, a Fab first chain-coding sequence or Fab second chain-coding sequence, and a scaffold-coding sequence in this order, the first Fab-expressing cistron further comprising at its 3-end a ribosome stall sequence, said Fab second chain-expressing cistron further comprising at its 3-end a structure necessary for maintaining a complex with the Fab encoded by itself.

The invention relates to a method and a prokaryotic cell for identifying a compound, which interferes with ribosome biogenesis, assembly and/or degradation. The cell expresses a first fusion protein comprising a first ribosomal protein, an amino acid linker and a first fluorescent protein, and a second fusion protein comprising a second ribosomal protein, an amino acid linker and a second fluorescent protein. The invention further relates to a gene construct comprising a first element encoding a first ribosomal protein fused to a first fluorescent protein by an amino acid linker, and a second element encoding a second ribosomal protein fused to a second fluorescent protein by an amino acid linker.

The present invention relates to methods of assembling a plurality of genetic units to form synthetic genetic constructs. This method involves appending universal adapter oligonucleotides and flexible adapter oligonucleotides to the 5 and 3 ends of separate genetic units to be assembled to form separate dual extended genetic units. The dual extended genetic units are assembled together via homologous recombination between the flexible adapter oligonucleotide portions of the dual extended units to form synthetic genetic constructs. The present invention further relates to synthetic genetic constructs formed using the methods of the present invention, and vectors, cells, and organisms containing such synthetic genetic constructs.

The present invention pertains to the field of protein engineering, and provides means for obtaining stable molecules that specifically bind to a target selected amongst a large variety of ligands families. In particular, the present invention provides methods for obtaining a molecule specifically binding to a target of interest, through a combinatorial mutation/selection approach with an OB-fold protein as a starting molecule. In particular, the target of interest can be of a different chemical nature form that of the native target of the OB-fold protein used as the starting molecule.

Methods are provided for, inter alia, detecting nucleic acid molecules resistant to degradation, such as a plurality of RNA molecules bound to a ribosome, using various technologies including deep sequencing.

The invention relates to a complex comprising a phage particle, said phage particle comprising (i) a polypeptide; (ii) a nucleic acid encoding the polypeptide of (i); (iii) a connector compound attached to said polypeptide wherein said connector compound is attached to the polypeptide by at least three discrete covalent bonds. The invention also relates to libraries, and to methods for making complexes and to methods of screening using same.

The invention relates to a complex comprising a phage particle, said phage particle comprising (i) a polypeptide; (ii) a nucleic acid encoding the polypeptide of (i); (iii) a connector compound attached to said polypeptide wherein said connector compound is attached to the polypeptide by at least three discrete covalent bonds. The invention also relates to libraries, and to methods for making complexes and to methods of screening using same.

The present invention relates to a library of particles, the library displaying a plurality of different T cell receptors (TCRs), wherein the plurality of TCRs may consist essentially of TCRs which may comprise an alpha chain variable domain from a natural repertoire and a beta chain variable domain from a natural repertoire, wherein the alpha chain variable domain may comprise a TRAV12-2 or a TRAV21 gene product and the beta chain variable domain may comprise a TRBV6 gene product.

An object of the present invention is to provide molecules that bind to a target molecule in a pH dependent manner and a screening method for selecting such molecules. Provided is a screening method for selecting peptides that bind to a target molecule at a first pH and do not bind thereto at a second pH, including a step of preparing a peptide library in which each peptide contains at least one special amino acid that undergoes a pH-dependent change in the charge of the side chain thereof, a step of bringing the peptide library into contact with the target molecule and incubating them under the first pH condition and selecting peptides that bind to the target molecule, and a step of selecting, from the peptides which have bound to the target molecule, peptides that do not bind to the target molecule under the second pH condition.