申请人:THE BOARD OF TRUSTEES OF THE UNIVERSITY OF ILLINOIS
公开号:US11198855B2
公开(公告)日:2021-12-14
Conformationally-constrained helicases having improved activity and strength are provided. Methods of making conformationally-constrained helicases having improved activity and strength are provided. Methods of using conformationally-constrained helicases having improved activity and strength are provided. The present invention is based on the discovery of novel modified helicases that show dramatically enhanced helicase activity and increased strength as compared to unmodified helicases. As described further herein, it has been surprisingly discovered that, by controlling the conformation of certain subdomains such that the helicase remains in a closed form (e.g., by covalently crosslinking the 2B domain to the 1A domain or the 1B domain in a Rep helicase), a highly active and strong form of the helicase is achieved.
提供了具有更高活性和强度的构象受限螺旋酶。提供了制造具有更高活性和强度的构象受限螺旋酶的方法。提供了使用具有更高活性和强度的构象受限螺旋酶的方法。本发明基于新型修饰螺旋酶的发现,与未经修饰的螺旋酶相比,这种螺旋酶的活性显著增强,强度也有所提高。如本文进一步描述的那样,令人惊奇地发现,通过控制某些亚结构域的构象,使螺旋酶保持封闭形式(例如,通过共价交联 2B 结构域与 Replicase 中的 1A 结构域或 1B 结构域),可以获得高活性和高强度形式的螺旋酶。