Cloning of the Gene Fragment Encoding the Coenzyme-binding Domain of Yeast ADH I and Its Fusion Expression in E. coli.

In this communication we report the cloning and the over-expression of the coenzyme-binding domain of yeast alcohol dehydrogenase. A gene fragment about 400 bp encoding the NAD binding domain of yeast S. cerevisiae alcohol dehydrogenase I was obtained by the PCR method. The domain was expressed as a fusion protein by a plasmid which was constructed by inserting the PCR-amplified gene fragment into the plasmid pWR590-1 which includes E. coli Lac promoter and a truncated Beta-galactosidase gene. The fusion protein was characterized by SDS-PAGE and Western blot analysis. The yield of the fusion protein was 20% of the total E. coli protein.