Recombinant DNA technology is a technique that alters the phenotype of an entity (host) when a genetically modified vector is introduced and incorporated into the genome of the host. Thus, the process entails introducing a foreign fragment of DNA into the genome containing the desired gene. This particular gene that is introduced is referred to as the recombinant gene and the technique is known as the recombinant DNA technology. Embedding the gene of interest into the genome of the host is not as simple as it sounds. Developing a recombinant DNA involves a series of sequential steps which are discussed below.
Process of Recombinant DNA Technology
Recombinant DNA technology involves the selection of the desired gene for administration into the host followed by a selection of the perfect vector with which the gene has to be integrated and hence the recombinant DNA is formed. This recombinant DNA, then has to be introduced into the host. And at last, it has to be maintained in the host and carried forward to the offspring.
Let’s go through the complete process in detail.
Isolation of DNA
Being a nucleic acid enclosed within the nucleus, isolation of DNA is not an easy task. Isolation of DNA is an enzymatically controlled process where the plant or animal cells are treated with certain enzymes. Enzymes such as cellulase (plant cells), lysozyme (bacteria) and chitinase (fungi) are used to isolate pure DNA from the cells.
Fragmentation of DNA
The isolated and purified DNA is treated with restriction endonucleases which cut the DNA into fragments. The restriction enzymes utilized in recombinant DNA technology are significant to detect the location at which the desired gene is introduced into the vector genome. The restriction endonucleases are sequence-specific, typically palindrome sequences and snip the DNA at specific points. They inspect the length of DNA and trims it at particular sites known as the restriction site. The desired genes and the vectors are snipped by the same restriction enzymes to acquire the complementary sticky ends. This ensures the task of ligases for binding the required gene to the vector is easier.
Amplification of Gene of Interest
Polymerase chain reaction (PCR) is a process to amplify the gene once the proper gene of interest has been cut using the restriction enzymes. Through this process, multiple copies of the gene of interest can be produced. PCR proceeds in three stages, denaturation, annealing and extension.
Insertion of recombinant DNA into the host
The host is the final tool of rDNA technology, which consumes the vector engineered with the desired DNA with the aid of the enzymes. Insertion of the desired recombinant DNA into the host organism can be achieved in various ways. This includes– biolistics or the gene gun, microinjection, alternate heating and cooling, usage of calcium ions, etc. The successfully transformed cells or the entities pass the recombinant gene to the offspring.
Stay tuned with BYJU’S to learn more in detail about the process of rDNA technology, its structure, and recombinant DNA technology.
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- Genetically Modified Organisms
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