Nour-Eldin Hussam H, Geu-Flores Fernando, Halkier Barbara A
Department of Plant Biology and Biotechnology, and VKR Centre Pro-Active Plants, Faculty of Life Sciences, University of Copenhagen, Frederiksberg, Denmark.
Methods Mol Biol. 2010;643:185-200. doi: 10.1007/978-1-60761-723-5_13.
The explosive development of the field of molecular biology has led to the need for simpler and more efficient cloning techniques. These requirements are elegantly met by the ligation-free cloning technique called USER cloning. USER cloning is suitable not only for everyday and high-throughput cloning but also for the one-step construction of complex DNA constructs, which can be achieved in a variant called USER fusion. In this chapter, we present a general protocol for converting any vector into a USER-compatible vector, together with protocols for both USER cloning and USER fusion.
分子生物学领域的迅猛发展使得对更简单、高效的克隆技术的需求日益凸显。名为USER克隆的无连接酶克隆技术出色地满足了这些要求。USER克隆不仅适用于日常和高通量克隆，还适用于复杂DNA构建体的一步法构建，这在一种称为USER融合的变体中能够实现。在本章中，我们将介绍把任何载体转化为USER兼容载体的通用方案，以及USER克隆和USER融合的方案。
