Expression plasmids containing the full cDNA sequence of calf prochymosin and capable of expressing prochymosin gene in E. coli host cells are disclosed. A method is described for the preparation of said plasmids which comprises altering the spacing between the SD and ATG sequences within the E. coli trp promoter-operator region of the parent expression plasmid, pCR 701, which is known to express prochymosin gene in the E. coli host cells. These modified expression plasmids are designed to provide high expression levels of prochymosin.