The document outlines the steps to engineer the insulin gene into E. coli cells which are: 1. Transforming the insulin gene and an antibiotic resistance gene into E. coli host cells 2. Accumulation of the beta-galactosidase/insulin fusion protein in the E. coli cells 3. Cleaving the insulin A and B chains from the fusion protein and purifying the chains 4. Mixing the purified A and B chains to form functional insulin

1. The sequence of steps to engineer the Insulin gene into E coli cells
Extract and purify ß-gal/ insulin
fusion proteins
Treat with cyanogen bromide to
cleave A and B chains
Purify, mix A and B chains to form
functional insulin
Disulfide bonds
Active insulin
ß galactosidase/ insulinA fusion
protein accumulatesin cell
ß galactosidase/ insulinA fusion
protein accumulatesin cell
Transform into E, coli host
Antibioticresistancegene
Insulin gene A subunit
ß gal genePromoter Promoter
ß gal gene
Insulin gene B subunit
Antibioticresistancegene

2. The sequence of steps to engineer the Insulin gene into E coli cells – With Labels Removed

3. The sequence of steps to engineer the Insulin gene into E coli cells – With Highlighted Part
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Antibiotic resistance gene

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