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Process for reconstituting the polymerase III* and other subassemblies of E. coli DNA polymerase III holoenzyme from peptide subunits
Process for reconstituting the polymerase III* and other subassemblies of E. coli DNA polymerase III holoenzyme from peptide subunits
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机译：从肽亚基重建大肠杆菌DNA聚合酶III全酶的聚合酶III *和其他子组件的方法
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摘要
The process of the invention provides for the reconstitution of the polymerase III* subassembly, Pol III*, of E. coli DNA polymerase III holoenzyme from substantially pure peptide subunits. In the first of two general schemes in which the subunits are added in a specified order, &ggr; and &tgr; are premixed before addition of &dgr; and &dgr;′. In the second general scheme, &dgr;′ is first assembled onto &ggr; (or &tgr;); then the excess &dgr;′ is removed before adding &tgr; (or &ggr;), following which &dgr; is added. Reconstituted Pol III* had the same subunit composition as purified natural Pol III*, as well as similar activity. Other smaller subassemblies of the polymerase III holoenzyme may also be reconstituted by the process of the invention.
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机译：本发明的方法提供了I E的聚合酶III *子组件Pol III *的重建。大体上纯肽亚基的大肠杆菌 DNA聚合酶III全酶。在以指定顺序添加子单元的两个通用方案中的第一个中，＆ggr;和＆tgr;在添加＆dgr;之前进行预混合和＆dgr;＆prime;。在第二种通用方案中，＆dgr;＆prime;首先组装到＆ggr;上（或＆tgr;）;然后是多余的＆dgr;＆prime;在添加＆tgr;之前已被删除（或＆ggr;），然后是＆dgr;被添加。重构的Pol III *具有与纯化的天然Pol III *相同的亚基组成，并且具有相似的活性。聚合酶III全酶的其他较小的子组件也可以通过本发明的方法重构。
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