Bioprocess Engineering 2nd Edition by Fikret Kargi,Michael Shuler

Exercise 4

You wish to produce a small protein using E. coli. You know the amino acid sequence of the
protein. The protein converts a colorless substrate into a blue product. You have access to a
high-copy-number plasmid with a penicillin-resistant gene and normal reagents for genetic
engineering. Describe how you would engineer E. coli to produce this protein. Consider:
source of donor DNA; regulatory elements that need to be included; how the donor and vec-
tor DNA are combined; how the vector DNA is inserted; and how you would select for a
genetically engineered cell to use in production.