Shuttling of entire libraries from an entry vector to a destination vector of the gateway system

Abstract

Large-scale experiments are the basis of functional genomics, the investigating of several hundred or even thousand genes or proteins in parallel. A prerequisite for such experiments is the ability to clone several thousand genes simultaneously into a vector of choice to investigate different aspects of protein function, e.g., protein interactions, or subcellular localization. In the recent past several such cloning systems have been developed and successfully used. Of the commercially available systems, the Gateway™ system is the most widely used. This protocol describes how to shuttle a library from an Entry vector to a destination vector of the Gateway™ system. Emphasis is placed on the efficiency of the shuttling process to avoid loss of complexity and on reproducibility of the method.