Co-Expression of Proteins in E. coli Using Dual Expression Vectors

AbstractDetailed biophysical, biochemical, and structural studies rely on the preparation of milligram amounts of pure recombinant proteins. Many useful overexpression systems have been developed for this purpose (1–3), and of these, bacterial overexpression is still the most convenient and simplest to use (4–6). Because many proteins are only active as heteromeric complexes, there has been much recent interest in preparing such complexes using recombinant technology (7–10). The preparation of complexes generally relies on the ability to reconstitute such protein complexes from individually prepared recombinant proteins (10–13), a process that often involves refolding (14,15). This is generally not ideal, and in cases where one protein is unstable without the other (16), impossible. KeywordsProtein ConstructUrea BufferNheI SiteInitial PurificationpRSET VectorThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.