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Staphylococcal nuclease fusion proteins for the production of recombinant peptides

机译:葡萄球菌核酸酶融合蛋白用于生产重组肽

摘要

Peptides are produced as fusions with a suitable carrier protein. The carrier protein disclosed herein are adapted from the N-terminal domain of staphylococcus nuclease. This novel carrier protein acts to promote the over-expression of the peptide-protein fusion in the form of inclusion bodies, which minimizes in-cell proteolysis of desired peptides. The fusion protein is readily purified by conventional procedures or His-tag affinity chromatography when His-tag is inserted into the fusion protein. The target peptide is released from the purified fusion protein by a simple cleavage step and separated from the librated carrier protein by use of a reverse-phase HPLC process or by repeating the same affinity purification method. A particular advantage of the disclosed method, in addition to the obvious advantage of high yields, is its use for producing isotopically labeled peptides for NMR characterization of bioactive peptides and their interactions with target proteins.
机译:肽与合适的载体蛋白融合产生。本文公开的载体蛋白质是从葡萄球菌核酸酶的N末端结构域改编的。这种新颖的载体蛋白以包涵体的形式促进肽-蛋白融合物的过表达,从而使所需肽的细胞内蛋白水解最小化。当将His-标签插入融合蛋白中时,可以通过常规方法或His-tag亲和色谱法容易地纯化融合蛋白。通过简单的切割步骤从纯化的融合蛋白中释放靶肽,并通过使用反相HPLC方法或重复相同的亲和纯化方法,从释放的载体蛋白中分离出目标肽。除了高产率的明显优点之外,所公开的方法的特别优点是其用于产生同位素标记的肽的用途,该肽用于NMR表征生物活性肽及其与靶蛋白的相互作用。

著录项

  • 公开/公告号US2009123972A1

    专利类型

  • 公开/公告日2009-05-14

    原文格式PDF

  • 申请/专利权人 ZHENGDING SU;FENG NI;

    申请/专利号US20070000370

  • 发明设计人 ZHENGDING SU;FENG NI;

    申请日2007-12-12

  • 分类号C12P21/04;C12N15/11;C12N15/00;C12N1/21;C12N1/19;

  • 国家 US

  • 入库时间 2022-08-21 19:35:10

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