Versatile, high-level expression cloning vehicles, pINIII, have been constructed. A foreign DNA fragment can be inserted in any one of the three reading frames at the unique EcoRI, HindIII, or BamHI site immediately after the initiation codon. The cloned foreign gene is under the control of both the lpp promoter and the lac promoter-operator. The expression of the gene is regulated by the lac repressor produced by the same vehicles. Using a pINIII vehicle EnvZ protein, a very minor regulatory protein of Escherichia coli was amplified more than 104-fold, becoming 30% of total cellular protein.
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