Gene cloning is a method in which a gene of interest is located and copied (cloned) out of all DNA extracted from an organism. It’s a consecutive method of making identical DNA or gene that is used in molecular biology. DNA is a polynucleotide chain and a tiny thing that can’t be seen without a microscope. Because of being small, it is tough to copy the DNA. In 1996, the first artificially cloned animal was born, is a sheep named “Dolly.” It has several downstream applications and is widely used in rDNA technology.
Steps of Gene Cloning:
Cloning a gene is a very sophisticated technique. Through some basic steps, researchers can clone a gene from a whole animal. These basic steps are-
1. Isolation of gene or DNA fragment: Firstly, the targeted gene should be isolated.
2. Suitable cloning vector selection: Then, you should choose the vector that can convey the gene of interest. Various vectors are available, like bacteriophages, plasmids, bacterial artificial chromosomes, etc.
3.Introducing recombinant DNA into the host cell: rDNA is introduced to the host through chemical, physical or enzymatic methods.
4.Using an antibiotic-resistant gene system, select the transformed host cell.
5.In the host cell, multiplication of the introduced gene.
6.It is isolating the copies of a gene.
7.Purifying the isolated gene.
Applications of Gene Cloning:
1.Gene cloning is used in gene and DNA analysis.
2.The presence and absence of a gene can be detected via gene cloning.
3.Specific genes can be isolated.
4.The nucleotide sequence of a gene can be fixed by using gene cloning.
5.Investigation of a protein/RNA function is also an application of gene cloning.
6.Mutation of a gene can also be detected by it.
7.The production of rDNA is one of the most important applications of gene cloning.
8.It is also used in pharmaceuticals.
Advantages of Gene Cloning:
Everything has its advantages and disadvantages. So cloning of a gene has also some advantages, for example-
1.Giving multiple copies of a gene or DNA is the prime advantage of gene cloning.
2.The solution to infertility.
3.It can provide organs for transplantation.
4.Offspring can be healthy.
5.The technique is less harmful.
Limitations of Gene Cloning:
Though gene cloning has many advantages, it also has some limitations. Like-
- Most of the techniques are time-consuming, but the gene cloning technique is not time-consuming. It takes more than 3 days to provide the result.
- It can be contaminated.
- Its accuracy is low.
- It losses the diversity of the gene.
- Religious problems.
After finding the polymerase chain reaction that is PCR, the gene cloning limitations were overcome in 1983. It is a cost-effective and rapid technique. It can give accurate and high performance. This can also provide more copies of genes than the conventional gene cloning technique. The main focus of the gene cloning technique is to multiply the gene or DNA. So it is the most powerful tool in molecular biology.
