Introduction
Recombinant DNA(rDNA) is a process that uses enzymes to attach the DNA sequences of interest. DNA sequences can be loaded into vectors, which transport the DNA to the appropriate host cell where it can be duplicated or reproduced. Human DNA, for example, may be programmed to replicate or reproduce bacteria or yeast. A bacterial cell or yeast to replicate this DNA next to it. This is known as DNA cloning and the result of compound DNA is sometimes called complex DNA. Find out what makes rDNA different and why it changes biology!
What exactly is rDNA Technology?
When a genetically modified vector is injected and integrated into the genome of an organism(host), the phenotypic of the organism changes. So, in essence, this process involves inserting an outer layer of DNA structure into a genome that encompasses our interesting genes. This injected gene is recurring, and the process is known as composite DNA technology. The DNA recombinant technique, used to produce synthetic DNA to produce the desired output, involves several stages, tools, and other processes.
Recombinant DNA Technology refers to the process of creating synthetic DNA by combining various genes(DNA) from different sources. Genetic engineering is another name for integrated DNA technology. Inserting the targeted gene into the host genome is not as easy as it seems. It involves selecting the gene needed to manage the host, followed by selecting the ideal vector at which genes need to be synthesized and reprogrammed DNA. The DNA recombinant must therefore be presented to the host. Finally, it has to be cared for locally and passed on to the offspring.
rDNA Technology is also known as Genetic Engineering
History of rDNA Technology
The emergence of DNA technology through the discovery of inhibitory enzymes in 1968 by Swiss biologist Werner Arber. In the early 1970s, the original DNA cells were reassembled and produced using inhibitory enzymes. Paul Berg, a Stanford scientist who was among the first to build a double DNA molecule in 1972, wrote the journal Science shortly thereafter, along with 11 other researchers.
Researchers from UC San Francisco and Stanford used blocking enzymes to break down DNA in a variety of ways in precise locations before assembling broken fibers back together. In 1974, Stanley Cohen of Stanford and Herbert Boyer of UCSF introduced patent DNA technology, which was granted in 1980.
Tools used in rDNA Technology
The restriction enzymes aid in cutting, the polymerases aid in synthesis, and the ligases aid in binding. The restriction enzymes employed in recombinant DNA technology are very important in identifying where the desired gene is introduced into the vector genome. The restriction endonucleases are sequence-specific enzymes that cut the DNA at specific locations. They are mainly palindrome sequences. They examine the length of the DNA and cut it at a specified location known as the restriction site. This causes the sequence to have sticky ends. The complementary sticky notes are obtained by cutting the appropriate genes and vectors with the same restriction enzymes, enabling the task of the ligases easier to bind the desired gene to the vector.
Aid in the transport and integration of the target gene. These are crucial components of recombinant DNA technology tools because they are the final vehicles for delivering the desired gene into the host organism. Plasmids and bacteriophages are the most commonly utilized vectors in recombinant DNA technology due to their high copy number.
Into which the recombinant DNA is inserted. The host is the ultimate instrument in recombinant DNA technology, taking in the vector built with the desired DNA with the assistance of enzymes. These chimeric DNAs are injected into the host in several ways, including microinjection, biolistics or gene gun, chilling and heating, calcium ion usage etc.
rDNA Technology Procedure
- Isolation of Genetic Material: The required DNA is extracted from other macromolecules in their pure form.
- The gene is cut at the recognition sites. The restriction enzymes play an important role in defining where the desired gene is introduced into the vector genome.
- The polymerase chain reaction is used to multiply gene copies (PCR). Once the appropriate gene of interest has been cut using restriction enzymes, it is a procedure that amplifies a single copy of DNA into hundreds to millions of copies.
- DNA Molecule Ligation: This involves the combining of two components–a cut segment of DNA and the vector–with the aid of the enzyme DNA ligase.
- Insertion of Recombinant DNA into the Host: The recombinant DNA is then injected into a recipient host cell in this stage. This is referred to as transformation. After the recombinant DNA is inserted into the host cell, it multiplies and is expressed in the form of the produced protein under ideal circumstances.
Technology shown in the bacterium Agrobacterium tumefaciens
Use-Cases of rDNA Technology
- rDNA technology is used to determine the existence of HIV in a person.
- Gene Therapy–It is used to try to fix the gene flaws that cause hereditary disorders.
- Clinical diagnostic–ELISA is an example of how recombinant DNA technology is utilized for medical diagnosis.
- It is used in agriculture to develop genetically-modified organisms such as Flavr Savr tomatoes, golden rice rich in proteins, Bt-cotton to protect the plant against ball worms and a variety of other products.
- Insulin is manufactured using Recombinant DNA technology in the pharmaceutical industry.
The development of rDNA technology resulted in a major wave of advancement and revolution in the fields of genetic engineering and gene cloning. It is a technology that serves as a foundation for the genetic industry.
Srishti Sharma
21-03-2022
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