METHOD FOR SELECTING (POLY) PEPTIDE/PROTEIN TAG CAPABLE OF COVALENTLY BONDING TO ANY TARGET SUBSTANCE AND (POLY) PEPTIDE/PROTEIN TAG SELECTED AND OBTAINED

• Main Authors: KAWAKAMI TAKASHI, NATSUME TORU
• Format: Patent
• Language: English; Japanese
• Published: 16.02.2017
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMBINATORIAL CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; LIBRARIES, e.g. CHEMICAL LIBRARIES, IN SILICOLIBRARIES; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; ORGANIC CHEMISTRY; PEPTIDES; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

PROBLEM TO BE SOLVED: To provide a method for obtaining at high speed a (poly) peptide/protein tag for protein-labeling, which is covalently bonded to a target substance.SOLUTION: The present invention provides a method for selecting a peptide or a protein, which is capable of covalently bonding to any target substance. The method comprises the steps of: (a) preparing an RNA library by transcribing a DNA library; (b) preparing a library of RNA-linker-nucleic acid derivatives; (c) performing a s peptide or protein synthesis in a cell-free translation system to produce an mRNA display peptide/protein library; (d) reverse-transcribing an mRNA part to DNA to prepare a cDNA display peptide/protein library; (e) contacting the cDNA display peptide/protein library with an immobilized target substance; and (f) selecting a conjugate in which a peptide portion or a protein portion of the cDNA display peptide/protein library is covalently bonded to the target substance.SELECTED DRAWING: Figure 1 【課題】標的物質と共有結合する、タンパク質標識用の（ポリ）ペプチド／タンパク質タグを、高速で得る方法の提供。【解決手段】以下の工程を含む、標的物質と共有結合可能なペプチド又はタンパク質の選択方法。(a)DNAライブラリーを転写してRNAライブラリーを作製する工程、(b)RNA−リンカー−核酸誘導体のライブラリーを作製する工程、(c)無細胞翻訳系でペプチド又はタンパク質合成を行い、mRNAディスプレイペプチド／タンパク質ライブラリーを作製する工程、(d)mRNA部をDNAに逆転写して、cDNAディスプレイペプチド／タンパク質ライブラリーを作製する工程、(e)cDNAディスプレイペプチド／タンパク質ライブラリーに、固定化された標的物質を接触させる工程、及び(f)cDNAディスプレイペプチド／タンパク質ライブラリーのペプチド部又はタンパク質部と標的物質とが共有結合した結合体を選択する工程【選択図】図１