Chemoenzymatic Synthesis of a Gene Control Region

Abstract

By the phosphitetriester approach we synthesized the fragments for constructing a synthetically idealized promoter (SIP). The total construction consists of the promoter, operator and Shine-Dalgarno (SD) region. This control region was fused to the gene for γ-interferon.

We chemically synthesized a control region in order to optimally express our synthetic genes considering the following criteria. For easy modifications, e.g. for structure function studies we designed each functional domain as a cassette which can conveniently be dissected by the appropriate restriction enzymes. The regulation unit consists of a promoter^1,2, an operator³ and a Shine-Dalgarno-sequence⁴ (FIG. 1).