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We have constructed a high-efficiency expression vector to direct the synthesis of heterologous polypeptides in yeast. The vector is termed a sandwich expression vector as the heterologous gene is inserted between the 5' and 3' control regions of the efficiently expressed yeast PGK gene. We have used this vector to direct the expression of three derivatives of the calf chymosin cDNA gene; preprochymosin, prochymosin and chymosin. Prochymosin is synthesised to at least 5% of total yeast-cell protein and furthermore, it can be readily activated to produce an enzyme which has milk-clotting activity.

Type

Journal article

Journal

Gene

Publication Date

09/1983

Volume

24

Pages

1 - 14

Keywords

Animals, Chymosin, Enzyme Activation, Genetic Vectors, Milk, Molecular Weight, Phosphoglycerate Kinase, Plasmids, Protein Precursors, RNA, Messenger, Saccharomyces cerevisiae