公开(公告)号：US06821755B2

公开(公告)日：2004-11-23

申请号：US09916229

申请日：2001-07-27

申请人： Rolf-Günther Werner ,  Klaus Bergemann ,  Friedrich Götz ,  Andreas Peschel

发明人： Rolf-Günther Werner ,  Klaus Bergemann ,  Friedrich Götz ,  Andreas Peschel

IPC分类号： C12P2102

CPC分类号： C12P21/02

摘要： The invention relates to advantageous processes for preparing heterologous proteins in prokaryotic host cells by improved codon use and/or expression of tRNAs which code for codons occurring rarely in said host cell.

摘要翻译： 本发明涉及通过改进密码子使用和/或表达在所述宿主细胞中很少发生密码子的tRNA编码的原核宿主细胞中制备异源蛋白质的有利方法。

公开(公告)号：US06566062B1

公开(公告)日：2003-05-20

申请号：US09508524

申请日：2000-05-02

申请人： Andreas Strauss ,  Gunther Thumm ,  Johannes Pohlner ,  Friedrich Götz

发明人： Andreas Strauss ,  Gunther Thumm ,  Johannes Pohlner ,  Friedrich Götz

IPC分类号： C12Q100

CPC分类号： C12Q1/02

摘要： A method for identifying a nucleic acid which codes for a polypeptide factor affecting the covalent bonding of polypeptides to the surface of Gram-positive bacteria, comprising the following steps: a) providing a sample of Gram-positive bacteria which can be genetically altered and contain or produce at least one enzymatic reporter substance which is or can become covalently bonded to the surface of the Gram-positive bacteria, said at least one reporter substance having a different enzymatic activity when not covalently bonded to the surface of the Gram-positive bacteria from that exhibited when it is covalently bonded to the surface of the Gram-positive bacteria; b) causing genetic alterations in Gram-positive bacteria of the sample; c) assaying the enzymatic activity of the reporter substance of the Gram-positive bacteria of the sample; d) separating Gram-positive bacteria which exhibit a different enzymatic activity of the reporter substance from that observed for covalent bonding of the reporter substance to the surface of the Gram-positive bacteria; e) isolating the nucleic acid of the Gram-positive bacteria separated in step d); f) identifying at least one segment of the nucleic acid isolated in step e) that carries the genetic alteration; g) based on the segment identified in step f), identifying a nucleic acid which codes for a polypeptide factor affecting the covalent bonding of polypeptides to the surface of Gram-positive bacteria.

摘要翻译： 一种用于鉴定编码影响多肽与革兰氏阳性细菌表面的共价键的多肽因子的核酸的方法，包括以下步骤：a）提供革兰氏阳性菌的样品，所述革兰氏阳性菌可被遗传改变并含有 或产生至少一种酶促报告物质，其是或可以共价键合到革兰氏阳性细菌的表面，所述至少一种报告物质在不共价结合到革兰氏阳性细菌的表面时具有不同的酶活性 当它与革兰氏阳性菌的表面共价结合时显示; b）引起样品的革兰氏阳性菌的遗传改变; c）测定样品革兰氏阳性菌的报道物质的酶活性 ; d）分离出与报道物质不同的酶活性的革兰氏阳性细菌与用于共价键合观察到的那些 e向所述革兰氏阳性细菌的表面提供记录物质; e）分离在步骤d）中分离的革兰氏阳性菌的核酸; f）鉴定在步骤e）中分离的核酸的至少一个片段，其携带 遗传改变; g）基于步骤f）中鉴定的区段，鉴定编码影响多肽与革兰氏阳性菌表面共价键的多肽因子的核酸。