Recombination DNA technology is of great importance in the field of medicine. With the help of a flow chart, show how this technology has been used in preparing genetically engineered human insulins.

Flowchart for Genetically Engineered Human Insulin Production using Recombinant DNA Technology:

1. Isolation of Human Insulin Gene:

   • Human insulin gene is isolated from human pancreatic cells using restriction enzymes.
   • This gene is then purified.

2. Vector Selection:

   • A suitable vector (e.g., plasmid) is chosen. Plasmids are small, circular DNA molecules that can replicate independently within bacterial cells.
   • The vector is chosen based on its ability to replicate efficiently and its compatibility with the host organism.

3. Gene Insertion (Recombination):

   • The isolated human insulin gene is inserted into the chosen vector using the same restriction enzyme used in step 1. This creates recombinant DNA.
   • DNA ligase enzyme joins the insulin gene and vector DNA, making a stable, recombinant plasmid.

4. Transformation:

   • The recombinant plasmid is introduced into a suitable host organism, typically E. coli bacteria, through a process called transformation.
   • This process involves making the bacterial cells competent to take up the foreign DNA.

5. Selection and Screening:

   • Transformed bacteria containing the recombinant plasmid are selected using a selective medium (e.g., one containing an antibiotic that only bacteria with the plasmid can survive in).
   • Bacteria that have successfully taken up the plasmid are further screened (e.g., using PCR or other molecular methods) to confirm that they carry the human insulin gene.

6. Production of Insulin:

   • The selected bacteria are cultured in large-scale fermenters. As they grow, they produce human insulin.

7. Purification:

   • The human insulin is extracted from the bacterial culture and purified using various techniques such as chromatography and filtration to remove bacterial proteins and other impurities.

8. Formulation and Packaging:

   • The purified insulin is formulated into a suitable form (e.g., solution or powder) for injection.
   • It's then packaged and tested for quality and safety before it is distributed.

Visual Flowchart (Text Representation):

[Human Pancreatic Cells] --> [Isolation of Human Insulin Gene] --> [Restriction Enzyme] --> [Purified Human Insulin Gene]
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[Vector (Plasmid)] --[Restriction Enzyme]-->(Recombinant DNA)
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[DNA Ligase] --> [Recombinant Plasmid]
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[E.coli Bacteria] --> [Transformation] --> [Transformed Bacteria (carrying recombinant plasmid)]
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[Selection and Screening] --> [Selected Bacteria]
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[Large-Scale Culture] --> [Insulin Production]
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[Purification] --> [Purified Insulin]
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[Formulation & Packaging] --> [Human Insulin Product]