Recombinant DNA technology can be used in various fields. Depending on where it is required, two DNAs of interest are combined and inserted into the host organism to produce a new genetic combination that can be used in the required area that can be medicine, science, agriculture, and industry. Recombinant DNA is the name for the DNA that is made by combining the other two DNAs.
There are certain laboratory methods for recombination to bring the genetic material together from the sources thus creating a different sequence that is new and unique. It was first achieved by Herbert Boyer in 1973 who inserted foreign DNA into the plasmids with the use of E.coli restriction enzymes.
The Steps of The Recombinant DNA Technology Process
Here are the steps that are followed in the process of making a recombinant DNA from two different DNAs:
1. Isolation of the Genetic Material/DNA
Once it is decided which DNAs are required to mix up, the very first step is to isolate that genetic material from the host and other macromolecules attached to it. Macromolecules like proteins, lipids, RNA are to be separated from the DNA. For this purpose, enzymes like cellulose, chitinase, proteases, etc. are used.
2. Restriction Enzyme Digestion
Restriction Enzymes are like the scissors that cut out the DNA from the specific parts. The purified DNA is put into incubation with the selected enzyme to make the conditions optimal for the enzyme to work.
It involves the use of polymerase chain reaction where there are multiple copies made of the DNA sequence with the help of an enzyme – DNA polymerase. It amplifies a single copy into thousands and millions of copies.
4. Ligation of DNA molecule
This is the process where the cut fragments of both the DNAs are joined to make the new DNA. The resulting DNA is the hybrid of the two DNA molecules and is called recombinant DNA.
5. Inserting the Recombinant DNA into the Host
Now that the hybrid/recombinant DNA is made, the next step is to transfer it into the host cell where the new DNA can grow and show its properties. Processes like thermal shock, Ca++ ion treatment, and electroporation are used to pursue he
6. Isolation of the Recombinant DNA
Once the DNA is transferred to the host, there is a mixed number of transformed and non-transformed cells. Now, the task is to separate and extract the transformed host cells that can grow and be useful. For this purpose, the marker gene is used which helps in isolating the host cell.
Uses of Recombinant DNA Technology
Here are some of the applications of recombinant DNA technology:
- It is widely used in medicine, research, and biotechnology.
- It can be used to identify the sequence of the genes to know more about it.
- These are widely used in laboratory experiments.
- This technology can also be used to identify and correct any genetic defect leading to hereditary diseases.
- It can also be used to identify HIV in a person.
- It also has wide applications in the agriculture department.
- It also helps the professionals in clinical diagnosis.
- It is also used in the process of application of medicine such as it is used to produce insulin.
Limitations of Recombinant DNA Technology
Where there are lots of uses of this technology, there are certain limitations too that make its use limited. Here are a few of them:
- Most professionals and scientists use it to destroy the native species for making a new one. It can disturb the balance of the environment and habitation.
- It contaminates the natural environment
- Many people have a religious point of view that it is wrong to play with God’s creation to make new species.
- Many people worry about hygiene and the safety of the food and medicines that are made through this technology.
- It can lead to people using stolen genetic information for their personal use of experimentation.
Where this technology has revolutionized the era, it has equally destroyed it by disturbing nature. However, its beneficial use in medicine can be considered helpful but once the experiments go out of the hands, it becomes unethical.