Recombinant DNA Technology in Today’s Medicine

Recombinant DNA (rDNA) technology involves combining DNA from different sources to create new genetic
sequences 1 . In simple terms, genes can be cut out of one organism’s DNA and inserted into another
organism’s DNA using molecular tools. This is possible because all DNA has the same chemical structure (A,
T, C, G bases) 1 . As a result, genes from humans, plants, bacteria, or viruses can be mixed. rDNA
technology is the foundation of modern biotechnology and medicine. It lets scientists mass-produce
important proteins and develop new therapies. For example, the insulin gene was inserted into bacteria
to produce human insulin in large amounts 2 , replacing the older method of extracting insulin from
animal pancreases.

Objectives
• To understand what recombinant DNA (rDNA) is and how it is created.
• To learn the key steps and principles of rDNA technology (e.g. cutting and joining DNA).
• To explore major applications of rDNA in medicine today.
• To examine examples of therapeutic proteins, vaccines, gene therapy, and monoclonal antibodies
made with rDNA.
• To review recent case studies showing the impact of rDNA in medical advances.

Principle and Process of rDNA Technology

The principle behind rDNA technology is simple: isolate a gene of interest and join it to a DNA carrier
(vector) so it can be replicated and expressed in a host organism. Scientists use restriction enzymes to cut
DNA at specific short sequences. These enzymes create “sticky ends” or blunt ends on both the gene and
the plasmid (a circular DNA vector) 3 . Then an enzyme called DNA ligase joins the cut ends to seal the
gene into the plasmid 4 . The result is a recombinant plasmid (vector + foreign gene) that carries the new
gene. This plasmid can be introduced (transformed) into bacteria or yeast. Inside the host cell, the plasmid
replicates and the inserted gene can be turned on to make the protein it encodes 4 .

The general steps in the process are:

1. Gene Isolation: Obtain the DNA segment (gene) of interest from any source (plant, animal, virus, etc.).
2. Restriction Digestion: Use restriction enzymes to cut the gene and a cloning vector (e.g. a bacterial
plasmid) at specific sites 3 . This produces compatible ends on both fragments.
3. Ligation: Use DNA ligase to join the gene and plasmid fragments into one circular DNA molecule,
forming recombinant DNA 4 .
4. Transformation: Introduce the recombinant plasmid into host cells (commonly E. coli bacteria) by a
process like heat shock or electroporation 5 .
5. Selection: Grow the transformed cells on antibiotic-containing media so only cells that have taken up the
plasmid (which carries an antibiotic resistance gene) will survive 5 6 . These surviving colonies carry and
express the foreign gene.
6. Expression and Harvest: Allow the host cells to multiply and express the recombinant gene, producing

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the desired protein (for example, insulin). Finally, the protein product is purified from the cells or their
culture medium.

Diagram: The illustration above outlines how a gene (red segment) is excised and inserted into a plasmid
vector (blue loop) using restriction enzymes and DNA ligase 3 4 . This recombinant plasmid is then
transformed into a bacterial cell to produce many copies of the new gene.

These tools (restriction enzymes, ligases, plasmids) allow precise “cut-and-paste” editing of DNA. The
plasmid vector is an autonomous circular DNA that can replicate in bacteria 7 . It usually contains marker
genes (like antibiotic resistance) and special sites (multiple cloning sites) that make insertion of foreign DNA
easier 5 . After ligation and transformation, scientists grow the bacteria on antibiotic plates. Only bacteria
containing the plasmid (and thus the new gene) survive and grow 5 6 . The figure below shows such a
laboratory plate experiment.

Illustration: Bacterial colonies grown on an antibiotic-containing petri dish. In this recombinant DNA
experiment, only bacteria that have successfully taken up the foreign gene (which includes an antibiotic-
resistance marker) can grow 6 . Colonies that fail to grow indicate bacteria lacking the plasmid, while
surviving colonies (circled) carry the new gene and can be studied further 6 5 .

Applications in Medicine
Recombinant DNA technology has revolutionized medicine. It enables the production of therapeutic
proteins, development of safer vaccines, creation of gene therapies, and manufacturing of monoclonal
antibodies, among other advances. Key applications include:

• Therapeutic Proteins: rDNA is used to mass-produce hormones and proteins that treat diseases.
For example, human insulin was the first biotechnology drug made this way 2 . Scientists inserted
the human insulin gene into E. coli, allowing bacteria to produce insulin identical to human insulin
2 . This recombinant insulin (approved in 1982) replaced animal-derived insulin, providing a pure

and unlimited supply. Similarly, the human growth hormone (hGH) gene was cloned so that
patients with growth hormone deficiency (GHD) could receive recombinant GH. The hGH gene was
first cloned in 1979, and a synthetic GH was introduced in 1985 8 9 . Recombinant proteins like
erythropoietin, interferons, clotting factors, and various enzymes are also produced by
engineered cells today. They are more consistent and safer than older methods.

• Vaccine Development: Many modern vaccines use rDNA technology. The Hepatitis B vaccine was
the first recombinant vaccine. It contains the Hepatitis B surface antigen (HBsAg) protein produced
in yeast cells. As described by McAleer et al. (1984), scientists inserted the HBsAg gene into yeast,
purified the antigen, and formulated it as a vaccine 10 . This was the first effective recombinant
vaccine against a human virus 10 . Other examples include recombinant subunit vaccines like the
HPV (human papillomavirus) vaccine, where the viral L1 protein is made in yeast. More recently,
COVID-19 vaccines have used novel rDNA-related approaches. Traditional viral-vectored vaccines
(like AstraZeneca’s) carry a coronavirus gene in a harmless virus, using rDNA to produce the antigen.
The new mRNA vaccines (Pfizer-BioNTech, Moderna) are based on in vitro-transcribed mRNA
encoding the viral spike protein. These mRNA instructions, designed using the virus’s gene
sequence, tell human cells to make the antigen 11 . Thus, even vaccine platforms rely on
recombinant genetic engineering to create safer and more effective immunizations.

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 • Gene Therapy: Gene therapy uses recombinant DNA to treat genetic diseases by delivering healthy
genes into patients. Typically, scientists attach a functional human gene to a viral vector (like AAV or
retrovirus) and administer it to the patient. For example, LUXTURNA (FDA-approved 2017) is an AAV-
based therapy that delivers a correct copy of the RPE65 gene to retinal cells, restoring vision in a
form of inherited blindness 12 . Zolgensma (approved 2019) uses AAV to deliver the SMN1 gene to
motor neurons, treating spinal muscular atrophy 12 . These gene therapies are built on rDNA
techniques to engineer the viral vectors with therapeutic genes. In total, dozens of gene therapy
products are approved or in trials, addressing cancer, rare diseases, and genetic disorders 12 .

• Monoclonal Antibodies (mAbs): Many modern drugs are monoclonal antibodies, and they are
produced by recombinant DNA methods. Recombinant mAbs are lab-engineered antibodies made by
inserting the antibody gene sequences into cultured cells 13 . The cells then secrete large amounts
of the antibody protein. For instance, Herceptin (trastuzumab) is a humanized antibody against
HER2 used in breast cancer, and Humira (adalimumab) targets TNF-alpha in autoimmune diseases.
These antibodies are produced by genetically modified mammalian cells (e.g. CHO cells) that carry
the cloned antibody genes. GenScript notes that recombinant mAbs offer greater design control and
consistency compared to older hybridoma methods 13 .

Table: Key medical applications of rDNA technology (examples and functions). The examples below illustrate
how specific genes are inserted into living cells to make useful products:

Application Example (Use) Role of rDNA Technology

Therapeutic Human insulin gene inserted into E. coli; bacteria

Insulin (for diabetes)
proteins produce insulin 2 .

Human growth hGH gene inserted into bacteria; produces

hormone (GHD) recombinant GH (approved 1985) 8 .

Vaccine HBsAg gene inserted into yeast; yeast produce viral

Hepatitis B vaccine
development antigen 10 .

COVID-19 mRNA SARS-CoV-2 spike protein gene used to make mRNA;

vaccines patient’s cells produce antigen 14 .

Luxturna (retinal AAV vector carries healthy RPE65 gene to eye cells
Gene therapy
disease) 12 .

Zolgensma (SMA) AAV vector delivers SMN1 gene to motor neurons 12 .

Monoclonal Herceptin (breast Antibody genes are cloned into cell lines; cells make
antibodies cancer) the antibody protein 13 .

Humira (rheumatoid Genes for anti-TNF antibody are expressed in cultured

arthritis) cells.

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Recent Examples and Case Studies
Modern breakthroughs show the power of rDNA in medicine. For example, the rapid development of
COVID-19 vaccines in 2020 used recombinant techniques: Pfizer/BioNTech and Moderna created synthetic
mRNA based on the coronavirus gene sequence, enabling a fast design and production pipeline. While this
is a new approach, it relies on gene-editing skills and synthesis of nucleic acids taught by rDNA methods
11 . Another example is CAR-T cell therapy for cancer, where a patient’s immune cells are genetically

modified to attack tumors (this involves inserting a chimeric receptor gene into T cells). In 2019, the
approval of Zolgensma for SMA patients was a landmark: an engineered AAV vector carrying the SMN1
gene was administered once and could halt disease progression 12 . These real-world cases illustrate how
rDNA technology is translating into patient cures and treatments in recent years.

Conclusion
Recombinant DNA technology has transformed modern medicine. By allowing precise gene splicing,
scientists can now manufacture proteins and develop therapies that were impossible before. rDNA has
given us effective hormones (insulin, GH), lifesaving vaccines (Hepatitis B, COVID-19), advanced cancer
treatments (mAbs and CAR-T), and cures for genetic diseases (gene therapy). It continues to be the engine
of biotech innovation. In summary, rDNA technology is a critical tool in medicine today, enabling
personalized treatments and improving health outcomes worldwide.

Acknowledgement
This project benefited from the guidance of my biology teacher and numerous online scientific resources. I
thank my teacher and peers for feedback, and I appreciate the open-access scientific publications and
educational websites that provided information. Special thanks to the creators of the diagrams and images
(all credited as public domain or Creative Commons).

References
• Quianzon C.C., Cheema A. (2013). History of Insulin. Journal of Community Hospital Internal Medicine
Perspectives. 2
• Ayyar V.S. (2011). History of Growth Hormone Therapy. Indian Journal of Endocrinology and
Metabolism. 8 9
• McAleer W.J. et al. (1984). Human hepatitis B vaccine from recombinant yeast. Nature 307, 178–180. 10
• Davidopoulou C., Kouvelas D., Ouranidis A. (2024). Comparing vaccine manufacturing technologies:
recombinant DNA vs in vitro transcribed mRNA. Sci. Rep. 14:21742. 11
• Shchaslyvyi A.Y. et al. (2023). Current State of Human Gene Therapy: Approved Products and Vectors.
Genes (MDPI). 12
• GenScript Biotech. (2023). Recombinant Monoclonal Antibody (mAb). 13 (accessed 2023)
• Suza W. (2010). Recombinant DNA Technology. LibreTexts Biology. 3 4
• Wikimedia Commons. Bacterial Transformation (diagram, CC BY-SA 3.0). 【58†】
• National Cancer Institute (1980). Recombinant DNA lab plate photo. Public domain. 6

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 1 Recombinant DNA - Wikipedia
https://en.wikipedia.org/wiki/Recombinant_DNA

2 History of insulin - PMC

https://pmc.ncbi.nlm.nih.gov/articles/PMC3714061/

3 4 5 1.11: Recombinant DNA Technology - Biology LibreTexts

https://bio.libretexts.org/Bookshelves/Genetics/Genetics_Agriculture_and_Biotechnology_(Suza_and_Lee)/01%3A_Chapters/
1.11%3A_Recombinant_DNA_Technology

6 File:Recombinant DNA.jpg - Wikimedia Commons

https://commons.wikimedia.org/wiki/File:Recombinant_DNA.jpg

7 Plasmid - Wikipedia
https://en.wikipedia.org/wiki/Plasmid

8 9 History of growth hormone therapy - PMC

https://pmc.ncbi.nlm.nih.gov/articles/PMC3183530/

10 Human hepatitis B vaccine from recombinant yeast - PubMed

https://pubmed.ncbi.nlm.nih.gov/6318124/

11 COMPARING vaccine manufacturing technologies recombinant DNA vs in vitro transcribed (IVT)

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mRNA | Scientific Reports

https://www.nature.com/articles/s41598-024-67797-x?error=cookies_not_supported&code=70a01c28-3ec6-4654-
b79d-8b65f647972f

12 Current State of Human Gene Therapy: Approved Products and Vectors - PMC
https://pmc.ncbi.nlm.nih.gov/articles/PMC10609992/

13 Recombinant monoclonal antibody - Terminology of Antibody Drug for Recombinant monoclonal

antibody – GenScript
https://www.genscript.com/biology-glossary/17595/recombinant-monoclonal-antibody