The recombinant gene is the gene that has been inserted, and the technique used to do so is referred to as recombinant DNA technology.
When it comes to recombinant DNA technology, there are numerous processes, instruments, and other particular procedures that must be followed to produce artificial DNA that will be used to create the intended product. Take a closer look at each of these steps in detail.
What is Recombinant DNA Technology and how does it work?
The term “Recombinant DNA Technology” refers to the process of creating artificial DNA by combining different genetic materials (DNA) from different sources. Recombinant DNA Technology is used to create artificial DNA through the combination of different genetic materials (DNA) from different sources. Genetic engineering is the term used to refer to the application of recombinant DNA technologies.
During the year 1968, Swiss microbiologist Werner Arber discovered restriction enzymes, which paved the way for the development of recombinant DNA technology.
It is not as simple as it appears to insert the desired gene into the genome of a living organism. Initially, it entails selecting the desired gene for administration into the host, followed by selecting the optimal vector into which the gene should be integrated and recombinant DNA produced.
As a result, it is necessary to introduce recombinant DNA into the host. Last but not least, it must be preserved in the host and passed on to the offspring to be effective.
Recombinant DNA Technology: Instruments for Success
In addition to restriction enzymes, polymerases and ligases are enzymes that aid in the synthesis of proteins and the binding of proteins to one another. The restriction enzymes that are used in recombinant DNA technology serve an important role in deciding where the desired gene is introduced into the vector genome, which is a crucial step in the process. There are two kinds of endonucleases: exonucleases and endonucleases.
The Endonucleases cut within the DNA strand, whereas the Exonucleases remove nucleotides from the ends of the DNA strands. Endonucleases are found in both bacteria and humans. The restriction endonucleases are sequence-specific enzymes that cut DNA at specified locations in the sequence, which is usually a palindrome sequence. They examine the length of the DNA and cut it at a specified point known as the restriction site to ensure that the DNA is not damaged. As a result, there are several sticky ends in the sequence. The desired genes and the vectors are both cut by the same restriction enzymes to produce the complementary sticky notes, which makes the work of the ligases to bind the chosen gene to the vector much easier to accomplish.
The vectors aid in the transportation and integration of the desired gene. These are a very significant component of the recombinant DNA technology toolset because they are the ultimate vehicles for delivering the desired gene into the host organism. Plasmids and bacteriophages are the most commonly utilised vectors in recombinant DNA technology because they have a high copy number and can be replicated indefinitely. The vectors are composed of three components: an origin of replication (which is a nucleotide sequence from which replication begins), a selectable marker (which is a gene that confers resistance to certain antibiotics such as ampicillin), and cloning sites (which are the sites recognised by restriction enzymes where desired DNAs are inserted).
The organism into which the recombinant DNA is injected is referred to as the host organism. The host is the most important tool in recombinant DNA technology since it is the only one that can take in the vector built with the desired DNA with the assistance of enzymes.
There are a variety of methods for introducing these recombinant DNAs into the host, including microinjection, biolistics or gene gun, alternate cooling and heating, the use of calcium ions, and other techniques.
Steps are the isolation of genetic material.
- The first and most important stage in the development of Recombinant DNA technology is to isolate the desired DNA in its pure form, that is, free of other macromolecules, from the surrounding environment.
- Cutting the gene where it binds to the recognition sites
It is the restriction enzymes that play a critical role in deciding where the desired gene is inserted into the vector genome, and they are very important. These kinds of reactions are referred to as “restriction enzyme digestions.”
Step three involves the use of a polymerase chain reaction to increase the number of gene copies (PCR).
Amplification of DNA is a procedure that is used to multiply a single copy of DNA into hundreds of millions of copies once the appropriate gene of interest has been cut using restriction enzymes.
Ligation of DNA Molecules is the fourth step.
Ligation is the process of connecting two pieces of DNA – a cut fragment of DNA and a vector – together with the aid of the enzyme DNA ligase during the replication process.
Insertion of recombinant DNA into the host cell (Step 5).
In this stage, the recombinant DNA is injected into a recipient host cell that has been selected. Transformation is the phrase used to describe this process. Once the recombinant DNA has been successfully inserted into the host cell, it can be replicated and expressed in the form of the produced protein under optimal conditions if the parameters are met.
Conclusion
Therefore it can be concluded, As previously noted in the section on Tools of recombinant DNA technology, there are a variety of approaches that can be used to accomplish this. The recombinant gene is passed on to the children by the successfully transformed cells/organisms.