Circular permutation is a powerful tool to test the role of topology in protein folding and function. Previous methods for generating circular permutants were based on rearranging gene elements using restriction enzymes-based cloning. Here, we present a Restriction Free (RF) approach to achieve circular permutation which is faster and more cost-effective.

中文翻译：

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使用无限制（RF）克隆促进循环排列。

循环置换是测试拓扑在蛋白质折叠和功能中的作用的强大工具。先前产生环状置换子的方法是基于使用限制酶克隆的基因元件重排。在这里，我们提出了一种无限制（RF）的方法来实现循环置换，该置换更快且更具成本效益。