Recombinant DNA Technology

• Recombinant DNA technology, also known as genetic engineering or gene cloning, is a set of techniques used to manipulate and modify DNA from different sources to create new DNA molecules.
• These recombinant DNA molecules are widely used in gene therapy, biotechnology, and pharmaceuticals.

Steps in Recombinant DNA Technology

1. Isolation of DNA

   • DNA is extracted from a source organism using enzymatic digestion or commercial extraction kits.

2. Cutting DNA

   • Restriction enzymes cut the DNA at specific sites, creating sticky or blunt ends that allow fragments to be joined with DNA from other sources.

3. Cloning Vectors

   • Vectors (e.g., plasmids or bacteriophages) carry foreign DNA into a host cell.
   • DNA fragments are inserted into vectors using sticky ends.

4. Transformation

   • The recombinant DNA is introduced into a host cell (e.g., bacteria) through transformation, allowing replication of the recombinant molecule.

5. Selection

   • Selection markers (e.g., antibiotic resistance genes) help identify host cells that successfully took up the recombinant DNA.

6. Screening

   • PCR, DNA sequencing, and Southern blotting verify the presence and integrity of the desired recombinant DNA.

Example: Production of Human Insulin

1. The insulin gene is isolated from human DNA.
2. It is inserted into a plasmid vector (e.g., pBR322).
3. The recombinant plasmid is introduced into E. coli.
4. The bacteria produce human insulin, which is later purified for medical use.

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