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Discuss how the cloning and expression of certain genes allows for massive production of the desired product.

DNA cloning is the process of making multiple, Identical versions of a specific piece of DNA A conventional DNA cloning technique begins by inserting the gene or other DNA fragment of interest (for example, a gene for a medically significant human protein) into a circular piece of DNA known as a plasmid. The injection is carried out using enzymes that “cut and paste” DNA, resulting in a molecule of recombinant DNA, or DNA formed from fragments from different sources.

• The recombinant plasmid is then inserted into bacteria. Bacteria containing the plasmid are chosen and grown. They repeat the plasmid and transfer it down to their descendants as they multiply, creating copies of the DNA it produces.
• What is the point of duplicating a DNA sequence in a plasmid? In certain cases, we need a large number of DNA copies to perform experiments or create new plasmids. In other instances, the DNA fragment encodes a valuable protein, and the bacteria serve as "factories" to produce the protein. The human insulin gene, for example, is found in E. coli bacteria to produce insulin, which is used by diabetics.

Steps of DNA cloning

• DNA cloning is used for a variety of applications. As an example, consider how DNA cloning can be used in bacteria to synthesize a protein (such as human insulin).

The basic steps are:

1. Cut open the plasmid and "paste" in the gene. . This method makes use of restriction enzymes (which cut DNA) and DNA ligase (which joins DNA).

Cutting and pasting DNA

• How can DNA fragments from various sources be linked together? Two types of enzymes are often used in this method: restriction enzymes and DNA ligase.  

2. Insert the plasmid into bacteria. Antibiotic selection may be used to classify the bacteria that absorbed the plasmid.

Bacterial transformation and selection

• In a mechanism known as transformation, plasmids and other DNA may be inserted into bacteria such as the innocuous E. coli found in laboratories. During transformation, specially prepared bacterial cells are subjected to a shock (such as high temperature) that induces them to accept foreign DNA.

3. Grow a large number of plasmid-carrying bacteria and use them as "factories" to produce the protein. Purify the protein after it has been extracted from the bacteria.

Protein production

• We can expand a large culture of plasmid-bearing bacteria until we've identified a bacterial colony with the correct plasmid. The bacteria are then given a chemical signal that instructs them to produce the target protein.

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