Characterization and Production of Protein Complexes by Co-expression in Escherichia coli
The functional units within cells are often macromolecular complexes rather than single species. Production of these complexes as assembled homogenous samples is a prerequisite for their biophysical and structural characterization and hence an understanding of their function in molecular terms. Co-expression in Escherichia coli has been used routinely to decipher the subunit composition, assembly, and production of whole protein complexes. Such complexes can then be used to reconstitute protein/nucleic acid complexes in vitro. In this chapter we present protocols for the widely utilized ACEMBL and pET-MCN/pET-MCP vector series which enable the rapid and automated co-expression of protein complexes in Escherichia coli.
Key wordsProtein complexes Co-expression Escherichia coli ACEMBL pET-MCN pET-MCP Cloning SLIC Plasmid concatenation Expression tests High throughput Automation
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