Heterologous Protein Expression in the Xenopus Oocyte

Abstract

The Xenopus oocyte is a specialized single cell of colossal size (>1 mm diameter) that is highly amenable for microinjection and a stalwart model for heterologous expression. Oocytes are easily obtainable, robust in vitro, and faithfully express injected constructs. Their large size translational capacity provides a huge canvas for observing and recording integrated cellular responses—from studies of single molecules within single cells to medium-throughput drug-screening applications. Most eukaryotic promoters suffice for Xenopus expression, and the oocyte can functionally express proteins from many diverse organisms. This protocol provides a basic introduction for scientists keen to perform nuclear microinjections of cDNA constructs. These are easy methods to master, do not require elaborate equipment, and make accessible a wonderful model cell system for studying signaling, transport, cell architecture, and protein function.