Recombinant DNA Technology

DNA is a collection of molecules that is in charge of transporting and passing genetic information from parents to offspring. DNA is the genetic material of a cell that carries information from generation to generation. It is essential for the survival of the cell. For the betterment of an individual scientists evolve new methods for the benefit or to cure diseases.

Recombinant DNA

It is a modified DNA molecule that includes genes from multiple sources, either through laboratory techniques or through genetic recombination.

The technology used for producing recombinant DNA is known as Recombinant DNA technology, which is commonly known as genetic engineering.

• Recombinant DNA is a piece of DNA that is made by the combination of at least two genes from two different sources.
• Recombinant DNA is possible because the DNA of all organisms share the same chemical structure but different nucleotide sequences within the overall identical structure. 
• Recombinant DNA is also known as chimeric DNA as this DNA can be formed by different species of organisms like mythical chimers.

Uses of Recombinant DNA

Agriculture

Recombinant DNA plays an important role in food production for the number of products made by plants.

• In the case of plants, recombinant DNA has been used to increase the resistance to viruses/pests in crops.
• Recombinant DNA has increased resilience in harsh environmental conditions.

Animals

Recombinant DNA plays an important role in animals. In the case of animals, bovine somatotropin (bST), is a hormone that is inserted bacterially in dairy cows to increase milk production. 

Vaccines 

Recombinant DNA is used in vaccines that involve the direct injection of genetic material into the human body. A plasmid or DNA loop from a foreign antigen that is the target of immunization makes up this genetic material. Our cells absorb the DNA once it is injected into our muscle tissue and begin generating the foreign proteins that are encoded in the plasmids. These proteins help the immunological reactions that our body has to the target antigen. 
DNA vaccines have the potential to be safer, cheaper, and longer lasting than current vaccination methods.

Treatment of Diseases

Numerous more illnesses and ailments can be treated with recombinant DNA.

• Patients with diabetes have benefited greatly from the manufacturing of insulin using recombinant DNA technology. The safest and purest versions of insulin on the market can now be produced by scientists in a manner that makes human insulin identical to pancreatic insulin.
• Patients of infertility have benefited greatly from the Recombinant human chorionic gonadotropin (r-hCG), recombinant human luteinizing hormone (r-hLH), and recombinant human follicle stimulating hormone (r-hFSH) are all hormones that help ovulation and follicular maturation operate properly, which is required for fertilization to be successful.

Tools of Recombinant DNA Technology

Among the enzymes that facilitate cutting, synthesis, and binding are restriction enzymes, polymerases, and ligases. 

The restriction enzymes used in recombinant DNA technology have a significant impact on where the intended gene is inserted into the vector genome. The two types are endonucleases and exonucleases.

• The exonucleases remove the nucleotides off the ends of the strands, whereas the endonucleases cut within the DNA strand. 
• The restriction endonucleases are sequence-specific and cut the DNA at predetermined locations. These sequences are typically palindrome sequences. 

They examine the DNA’s overall length and remove a particular region known as a restriction site. As a result, the sequence develops sticky ends. The same restriction enzymes are used to cut both the vectors and the desired genes, resulting in complementary sticky notes. 

This makes it simple for the ligases to bind the desired gene to the vector. 

The desired gene is carried by and integrated into the vectors. They are the primary means by which the desired gene is transferred into the host organism, making them a crucial component of the recombinant DNA technology’s instruments. The most popular vectors employed in recombinant DNA technology are bacteriophages and plasmids because of their high copy numbers. The components of the vectors are the origin of replication, which is a sequence of nucleotides from which the replication begins, a selectable marker, which consists of genes that exhibit resistance to specific antibiotics like ampicillin, and cloning sites, which are the locations where desired DNAs are inserted and are recognized by restriction enzymes.

The organism is used as the host, into which the recombinant DNA is inserted. The host, which accepts the vector created with the required DNA with the aid of enzymes, is the ultimate tool of recombinant DNA technology. These recombinant DNAs can be introduced into the host in several ways, including microinjection, biolistics or gene gun, alternate cooling, and heating, usage of calcium ions, etc.

Process of Recombinant DNA Technology

Recombinant DNA technology involves several stages to yield the desired result.

Step 1: Isolate the genetic material

The primary stage in the creation of recombinant DNA is the isolation of the desired DNA in its purest form. It doesn’t contain any other macromolecules.

Step 2: Cutting the gene at the locations of recognition

The desired gene is inserted into the vector genome at that site after the restriction enzymes play a significant role in determining the location.” Restriction enzyme digestion.” This is the name of the procedure.

Step 3: Using the Polymerase Chain Reaction to increase the gene copies (PCR)

In this procedure, a single DNA copy is multiplied by hundreds to millions, and then the correct gene of interest is cut using restriction enzymes.

Step 4: Ligation of DNA molecules

With the aid of the enzyme DNA ligase, a DNA fragment and the vector are joined together in this phase.

Step 5: Recombinant DNA is inserted into the host

Transformation is a term used to describe the process of inserting recombinant DNA. Recombinant DNA is injected into a host cell using this technique. Recombinant DNA multiplies after being inserted into the host cell and is exposed to the produced protein form under optimum conditions. The recombinant gene is passed on to the progeny by the altered cells.

Application 

1. Biotechnology, medicine, and research all make extensive use of them.
2. It is used to make recombinant therapeutics like human insulin, interferon, and growth hormone as well as vaccines.
3. It is utilized in agriculture to create crops that are resistant to insects.
4. It helps with the creation of gene therapy.
5. It is used for hepatitis diagnostic tests and to find out if someone has HIV.
6. It’s utilized in the creation of transgenic animals.
7. It is applied in tests to identify paternity and hereditary diseases.
8. It produces clotting factors that are used to treat hemophilia.

DNA Cloning

Making a genetically identical replica of an organism is cloning. Individual cells, entire species, or even single genes can all be cloned. Making an identical copy of DNA is known as DNA cloning. DNA cloning is frequently employed in medical research to investigate disease states and develop new medications.

Applications

Gene cloning has several applications in the fields of medicine and science. Examples include:

• Biopharmaceuticals: Human proteins with biological uses are produced through DNA cloning. E. coli-based insulin synthesis is one instance. Other examples are tissue plasminogen activator, which is used to treat strokes, and human growth hormones, which are used to treat people who cannot naturally make this hormone.
• Gene therapy: Gene therapy is used to provide a patient with a specific gene that is missing from their body as a result of a genetic condition. It was possible to create plasmids with a normal copy of the cystic fibrosis gene via DNA cloning. Lung function decline was less rapid in cystic fibrosis patients after the plasmids were administered to their lungs.
• Gene Analysis: To understand the function of a particular gene and how it works in an organism, biologists frequently utilize DNA cloning to produce artificial, recombinant versions of genes.

Properties of Organisms containing Recombinant DNA

Recombinant DNA-containing creatures typically have characteristics that seem typical. As a result, the only way to prove the presence of recombinant sequences is to study the DNA itself, commonly through the use of a polymerase chain reaction (PCR) test. This means that their appearance, behavior, and metabolism are typically unaltered.

Gene Expression 

• Gene expression or DNA expression is the process by which our DNA’s instructions are transformed into useful products, such as proteins.
• A finely controlled process called gene expression enables a cell to react to its changing surroundings.
• It functions as a volume control that raises or lowers the production of proteins, as well as an on/off, switch for controlling when proteins are generated.
• Transcription and translation are the two essential processes that are involved in the production of a protein.
• The process of creating an RNA strand from a DNA strand is called transcription. DNA is once again used as a template during transcription, just as it is during DNA replication. A new RNA molecule is “transcribed” with the information previously contained in DNA molecules.
• The process of translation uses the data stored in mRNA molecules to produce proteins. A protein with a specified sequence of amino acids can be produced using the nucleotide sequence found in the mRNA molecule. The information contained in the strand of RNA is “translated” during translation from RNA language into polypeptide language, or the sequence of nucleotides into the sequence of amino acids.

Summary of rDNA

Recombinant DNA is a type of DNA that is created in a lab. It is created by moving certain DNA segments from one organism to another. There are numerous uses for DNA cloning. The mapping of the human genome, the generation of transgenic animals, and the construction of insect-resistant crops have all been shown to be extremely helpful. It is essential to genetic tests used in forensic research, and archaeology, as well as those used to establish paternity and diagnose hereditary diseases. Diagnostic tests for hepatitis and the human immunodeficiency virus (HIV) are also built using this technique. Vaccines and protein medicines like human insulin, interferon, and growth hormone are also produced using recombinant DNA technology. Additionally, it is employed in the creation of gene therapy and the production of clotting factors for the treatment of hemophilia.

FAQs on Recombinant DNA

Question 1: Define DNA and R-DNA.

Answer:

• DNA is a collection of molecules that is in charge of transporting and passing genetic information from parents to offspring.
• Recombinant DNA is a modified DNA molecule that includes genes from multiple sources, either through laboratory techniques or through genetic recombination.

Question 2: Why recombinant DNA is called chimeric DNA?

Answer:

Recombinant DNA is also known as chimeric DNA as this DNA can be formed by different species of organisms like mythical chimers.

Question 3: What is the use of recombinant DNA in agriculture?

Answer:

Recombinant DNA plays an important role in food production for the number of products made by plants.

• In the case of plants, recombinant DNA has been used to increase the resistance to viruses/pests in crops.
• Recombinant DNA has increased resilience in harsh environmental conditions.

Question 4: How does recombinant DNA help to treat diseases?

Answer:

Some numerous more illnesses and ailments can be treated with recombinant DNA.

• Patients with diabetes have benefited greatly from the manufacturing of insulin using recombinant DNA technology. The safest and purest versions of insulin on the market can now be produced by scientists in a manner that makes human insulin identical to pancreatic insulin.
• Patients of infertility have benefited greatly from the Recombinant human chorionic gonadotropin (r-hCG), recombinant human luteinizing hormone (r-hLH), and recombinant human follicle stimulating hormone (r-hFSH) are all hormones that help ovulation and follicular maturation operate properly, which is required for fertilization to be successful.

Question 5: Write a brief note on recombinant DNA.

Answer:

Recombinant DNA is a modified DNA molecule that includes genes from multiple sources, either through laboratory techniques or through genetic recombination. The technology used for producing recombinant DNA is known as Recombinant DNA technology, which is commonly known as genetic engineering. Recombinant DNA is a piece of DNA that is made by the combination of at least two genes from two different sources. Recombinant DNA is possible because the DNA of all organisms share the same chemical structure but different nucleotide sequences within the overall identical structure.  Recombinant DNA is also known as chimeric DNA as this DNA can be formed by different species of organisms like mythical chimer