Recombinant DNA Technology: Gene Cloning

• Recombinant DNA Technology modifies and manipulates an organism's genetic material.
• This biotechnology increases genetic variation by creating new genetic combinations.
• It introduces traits not previously in the gene pool.
• This produces recombinant DNA, which contains genes from two or more sources.
• Gene cloning using recombinant DNA technology involves four main steps.

Isolation

• A plasmid is isolated and extracted from an E. coli bacterium.
• A gene of interest is isolated and extracted from the DNA of an organism.

Digestion/Cutting

• The plasmid and the gene of interest are cut with the same restriction enzyme.
• Restriction enzymes act as scissors and create matching sticky ends.
• Sticky ends allow the fragments to fit together.

Insertion

• The gene of interest is inserted into the plasmid.
• This happens by aligning the sticky ends of both DNA fragments.

Ligation

• DNA ligase joins the gene and plasmid together
• This forms recombinant DNA.
• Recombinant DNA technology can develop transgenic organisms for agricultural and medical applications.
• It is used to put the human insulin gene into bacteria.
• The bacteria then produces the same insulin that humans would create.

Insulin Production Steps

• The human insulin gene and bacterial plasmid are isolated.
• Both are cut with the same restriction enzyme to create sticky ends.
• The insulin gene is inserted into the plasmid.
• DNA ligase joins the gene and plasmid.
• The recombinant DNA is inserted into bacteria to produce insulin.
• The insulin is extracted and purified in a lab.
• It's stored in vials or pens for treating diabetes.