Clone the human insulin gene for expression in E.coliBL21. Look at the DNA and mRNA sequence of insulin. The insulin gene will be under the control of a lacI repressor. Use these sequences along with plasmid pET11c to develop a method to express the human insulin gene in E. coli.
1. Explain how you would clone the insulin gene into the Hind III restriction sire of pET11c. Also how would you induce expression?
2. Determine how many orientations can the insulin gene be cloned intopET11c?
3. Explain how would you confirm the presence of your plasmid?