MOLL, GERT NIKOLAAS,LEENHOUTS, CORNELIS JOHANNES,KUIPERS, OSCAR PAUL,DRIESSEN, ARNOLD JACOB MATHIEU
申请号：
ES03733622
公开号：
ES2444840T3
申请日:
2003.05.26
申请国别(地区):
ES
年份:
2014
代理人:
摘要：
A method for producing in a host cell, a polypeptide containing a thioether bridge, whose origenno is a Gram-positive bacterium, the method comprising a) selecting a host cell having a nucleic acid molecule comprising a first and unsegundo nucleic acid fragment which are in the same open reading frame, wherein - the first nucleic acid fragment encoding a leader peptide lantibiotic that funcionalmenteequivalente is a leader peptide N-terminal prepeptide found in a lantibiotic, and wherein estepéptido acts as a signal leader sequence translocation and an acknowledgment signal, such maneraque forming a thioether linkage may occur - the second nucleic acid fragment encoding a desired sequence comprising a thioether bridge AModify a polypeptide sequence selected from the group of sequences consisting of Ser and Thr-Xaa-Cys-Xaa-Cys, where Xaa is any amino acid and wherein n is 1-13, and where polipéptidono has its origin in a bacterial cell Gram-positive b) selecting the host cell for the presence of the carrier protein LanT c) translating the nucleic acid molecule, thereby producing a fusion peptide of delantibiótico leader peptide and the polypeptide comprising the sequence selected from the group consisting of sequences porSer-Xaa-Cys-Xaa-Thr and Cys, where Xaa is any amino acid and wherein n is 1-13 d) harvesting the polypeptide containing the thioether bridge medium of the host cell.Un método para producir, en una célula hospedadora, un polipéptido que contiene un puente tioéter, cuyo origenno es una bacteria Gram-positiva, comprendiendo el procedimiento a) seleccionar una célula hospedadora que tenga una molécula de ácido nucleico que comprenda un primer y unsegundo fragmento de ácido nucleico que estén en la misma fase de lectura abierta, en la que - el primer fragmento de ácido nucleico codifica un péptido líder de lantibiótico que es funcionalmenteequivalente a un péptido líder N-terminal encontrado en un prepéptido de un lantibiótico, y en el que estepéptido líd
