• 主题
高级搜索  >
历史搜索 清空历史
首页> 外国专利> Vector for the heterologous protein expression and method for extraction of recombinant proteins as well as for isolating recombinant insulin

Vector for the heterologous protein expression and method for extraction of recombinant proteins as well as for isolating recombinant insulin

机译：用于异源蛋白表达的载体，重组蛋白的提取以及分离重组胰岛素的方法

页面导航
  • 摘要
  • 著录项
  • 相似文献

摘要

The present invention relates to a vector for expression of a heterologous protein by a Gram negative bacteria, wherein the vector includes a nucleic acid such as DNA encoding the following: an origin of replication region; optionally and preferably a selection marker; a promoter; an initiation region such as translation initiation region and/or a ribosome binding site, at least one restriction site for insertion of heterologous nucleic acid, e.g. DNA, encoding the heterologous protein, and a transcription terminator. The inventive vector may contain DNA encoding the heterologous protein, e.g., pro-insulin such as pro-insulin with a His tag. Additionally, the invention provides a method for extracting a recombinant protein from within a recombinant Gram negative bacteria having a cell membrane, without lysing the bacteria, as well as a method for purifying an isolated recombinant human insulin, wherein the isolated recombinant human pro-insulin is subjected to sulfitolysis, Ni-chelation chromatography, renaturation, limited proteolysis and chromatography separation to provide purified, isolated, recombinant human insulin.
机译：本发明涉及一种用于革兰氏阴性细菌表达异源蛋白的载体，其中该载体包括一种核酸，例如编码以下内容的DNA：复制起点区;任选地并且优选地，选择标记;发起人;起始区，例如翻译起始区和/或核糖体结合位点，至少一个用于插入异源核酸的限制位点，例如，RNA。编码异源蛋白质的DNA和转录终止子。本发明的载体可以含有编码异源蛋白质的DNA，例如具有His标签的胰岛素原，例如胰岛素原。另外，本发明提供了从具有细胞膜的重组革兰氏阴性细菌内提取重组蛋白而不裂解细菌的方法，以及纯化分离的重组人胰岛素的方法，其中分离的重组人胰岛素原将其进行亚硫酸酯化，Ni螯合色谱，复性，有限的蛋白水解和色谱分离，以提供纯化，分离的重组人胰岛素。

著录项

相似文献

  • 专利
  • 外文文献
  • 中文文献
获取专利

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号