Answer to Question #166771 in Genetics for Hafiz Faizal

Question

What THE PROCEDURES IN RECOMBINANT DNA TECHNOLOGY

and LIST THE APPLICATIONS AND DISADVANTAGES OF THIS TECHNOLOGY

Answer 1

Recombinant DNA technology is a series of procedures used to join together (recombine) DNA segments from different species. A recombinant DNA molecule is constructed from segments of two or more different DNA molecules that have been obtained from different sources.

The procedures involve;

• Treating the foreign DNA and plasmid with restriction enzymes and DNA ligase to make recombinant plasmid.
• Introducing the recombinant plasmid into cultured cells.
• Regenerating new organisms from the cultured cells

The complete process of recombinant DNA technology includes multiple steps, maintained in a specific sequence to generate the desired product.

Step-1. Isolation of Genetic Material.

The first and the initial step in Recombinant DNA technology is to isolate the desired DNA in its pure form i.e. free from other macromolecules.

Step-2.Cutting the gene at the recognition sites.

The restriction enzymes play a major role in determining the location at which the desired gene is inserted into the vector genome. These reactions are called ‘restriction enzyme digestions’.

Step-3. Amplifying the gene copies through Polymerase chain reaction (PCR).

It is a process to amplify a single copy of DNA into thousands to millions of copies once the proper gene of interest has been cut using the restriction enzymes.

Step-4. Ligation of DNA Molecules.

In this step of Ligation, joining of the two pieces – a cut fragment of DNA and the vector together with the help of the enzyme DNA ligase.

Step-5. Insertion of Recombinant DNA Into Host.

In this step, the recombinant DNA is introduced into a recipient host cell. This process is termed as Transformation. Once after the insertion of the recombinant DNA into the host cell, it gets multiplied and is expressed in the form of the manufactured protein under optimal conditions.

Application of Recombinant DNA Technology

• DNA technology used to detect the presence of HIV in a person.
• Gene Therapy - to correct gene defects which give rise to heredity diseases
• Clinical diagnosis e.g. ELISA
• In Agriculture to produce genetically-modified organisms.
• In medicine e.g. in the production of Insulin.

Disadvantages of Recombinant DNA technology

• Destruction of native species in the environment the genetically modified species are introduced in.
• Resilient plants can theoretically give rise to resilient weeds which can be difficult to control.
• Cross contamination and migration of proprietary DNA between organisms.