| 摘要 |
<p>A high throughput screening system showing a high efficiency for searching for a ligand of a target protein, in which a display random peptide library is used and which is applicable to a membrane protein, is constructed. The display random peptide library to be used therefor is a library consisting of gram-negative bacteria having been transformed by expression vectors, in which the individual polynucleotides in a random polynucleotide library comprising polynucleotides, encoding a target protein and having a secretion signal sequence located upstream, are connected in tandem; wherein said target protein is expressed on the inner membrane surface of each of the gram-negative bacteria; and each of polypeptides corresponding respectively to the polynucleotides constituting said random polynucleotide library is present in the periplasmic space. A polypeptide having bonded to the target protein is obtained by screening and recovering spheroplasts. The obtained polynucleotide is amplified and subjected, as the starting material in the subsequent cycle, to repeated screening and amplification so that the in vitro molecule evolution thereof can be conducted to thereby give a polypeptide sequence having a higher affinity for the target protein.</p> |
