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Question 6
a. Outline how human insulin is made using recombinant plasmids. There are different types of insulin available for use. The activity profile of each of these insul... show full transcript
Step 1
Answer
Human insulin is produced using recombinant DNA technology, which involves several crucial steps:
Gene Isolation: The genes for insulin chains A and B are isolated from human DNA using restriction endonucleases. These enzymes cut the DNA at specific sequences, allowing the desired genes to be extracted.
Plasmid Preparation: A plasmid, which is a small, circular piece of DNA, is prepared to carry the insulin genes. The plasmid is cut open to allow the insertion of the isolated insulin genes.
Insertion of Insulin Genes: The insulin genes are inserted into the plasmid using DNA ligase, which joins the DNA sequences together, creating a recombinant plasmid.
Transformation into Bacteria: The plasmid is introduced into a bacterial host through a process known as transformation. This allows the bacteria to take up the plasmid and start producing the insulin chains.
Selection and Culturing: Antibiotic selection or another method, such as inserting markers, is used to identify successfully transformed bacteria. The selected bacteria are then cultured to produce insulin.
Protein Processing: Finally, the insulin chains produced in the bacteria are harvested and processed. They are linked together to form functional insulin through techniques such as joining the polypeptide chains A and B.
Step 2
Answer
The type of insulin that is rapid acting compared to regular insulin is Lispro. From the graph, Lispro shows a steep increase in activity, peaking quickly at around 2 hours, with a maximum activity of approximately 5 to 7 arbitrary units. In contrast, regular insulin peaks later, around 5 hours, with a lower maximum activity. This indicates that Lispro acts more quickly, making it suitable for rapid insulin needs.
Step 3
Step 4
Answer
When altering the primary structure of insulin, scientists must consider the potential impact on the protein's tertiary and quaternary structures, as these levels are critical for its functionality. Any changes in the sequence of amino acids could lead to a misfolded protein, which might result in loss of biological activity or unintended side effects. Therefore, ensuring that alterations do not disrupt the functional properties of insulin is essential.
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