A plasmid was constructed that encodes all five subunits of the Escherichia coli -complex on a single artificially constructed operon under the control of an inducible promoter. The proteins , , `, , and overproduced from this artificial operon assemble efficiently in vivo, providing an efficient source of homogeneous -complex. The subunit is a truncated form of that is produced by a translational frameshift. When protein expression was induced in bacterial strains containing the outer membrane protein T (OmpT) protease, was proteolyzed after lysis to a -like protein, , and a peptide, C-, corresponding to the C terminus of . N-terminal sequencing of C- revealed a cleavage site between two lysines at positions 429 and 430 of . The deduced sequence of is, therefore, only two amino acids shorter than natural . The proteolysis by OmpT was also shown directly by using purified OmpT and -complex in an in vitro reaction. A -complex and a mixed --complex were purified from ompT cells. When the -complex proteins were overexpressed in ompT bacteria, intact -complex lacking could be purified.

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