Recombinant DNA
Technology
Group 5
At the end of this module, I can:
1. Outline the processes involved in genetic
engineering.
2. Discuss the applications of recombinant DNA.
3. State the importance of recombinant DNA.
Genetic engineering Modifies an organism's genetic
code to add new traits not naturally present. Advanced
DNA understanding allows scientists to extract, cut, and
copy DNA, enabling targeted genetic changes. DNA
recombination A process of modifying the genes of
organisms for practical purposes. It is done when a piece
of DNA is combined with another DNA from another
source. The resulting genetic product is called
recombinant DNA. With this process, organisms get to
have traits that are not normally found in their species.
Brief History of Recombinant DNA Technology In the late 1960s,
Stewart Linn and Werner Arber discovered restriction enzymes in E.
coli, which cut DNA at specific sites and create sticky ends for
joining DNA fragments. This foundational knowledge paved the
way for Herbert Boyer and Stanley Cohen's groundbreaking
experiments in 1973, where they successfully recombined plasmids
in E. coli, introducing genes that provided antibiotic resistance and
allowing the expression of frog DNA in bacteria.
The success of these experiments revolutionized genetic
engineering, enabling the manipulation of genes to
create organisms with desirable traits. However, by 1974,
growing public concerns about the ethical implications of
gene manipulation led ten scientists, including Paul Berg,
to express their worries in a letter to the National
Academy of Sciences. This prompted the Asilomar
Conference in February 1975, which aimed to establish
safety guidelines for the use of recombinant DNA
technology.
Technology and
Tools Used in
Recombinant DNA •Gel electrophoresis
Technology -Separates DNA fragments by
size. Negatively charged DNA
moves through a gel towards
a positive electrode; smaller
fragments move faster. This
technique is crucial for DNA
fingerprinting, genome
comparison, and gene
identification.
•DNA Sequencing
-This is a method used to provide the
identity and order of nucleotides in
a DNA strand. Small, single-stranded
pieces of DNA are placed in test
tubes with an enzyme that can
make a complementary DNA strand
by using the original DNA strand as a
template. A supply of the four
nucleotide bases found in DNA is
then added, along with a small
amount of one of the bases that has
been labeled with fluorescent dyes.
•Polymerase Chain Reaction
(PCR)
- Is a method used to amplify
specific DNA sequences,
essential for detecting diseases
and infectious agents. The
process involves heating DNA to
separate its strands, cooling to
allow primers to bind, and using
DNA polymerase to synthesize
new strands. This cycle of heating
and cooling is repeated, resulting
in millions of copies of the target
DNA. PCR is a vital tool in
molecular biology and
diagnostics.
Processes Used in
Recombinant
DNA Technology
•Transformation
-Using a Vector Recombinant DNA is created through
transformation using vectors like plasmids found in
bacteria. In this process, foreign DNA is inserted into a
plasmid, which serves as a carrier. A restriction enzyme
cuts the donor DNA to create sticky ends, allowing it to
pair with the plasmid. DNA ligase then seals the DNA
together. The plasmid contains a genetic marker for
identification, often an antibiotic resistance gene. After
transformation, bacteria are treated with antibiotics, and
only those that have taken up the plasmid survive,
allowing for the selection of successfully transformed cells.
Vectorless Gene Transfer
-Vectorless gene transfer introduces
foreign DNA into host cells without using
vectors. Techniques include
electroporation, protoplast fusion,
microinjection, and particle guns. In
electroporation, electrical pulses create
temporary holes in the cell membrane,
allowing DNA to enter. Protoplast fusion
involves digesting bacterial cell walls to
form protoplasts, which are then fused
using polyethylene glycol to mix their
genes. The resulting recombinant cells will
grow a new cell wall.
In microinjection, a host cell is held
in place using mild suction, and a
foreign gene is injected directly into
the cell with a fine needle, creating
recombinant DNA. The particle gun
method involves bombarding the
host cell with tungsten particles
coated with foreign DNA. This
technique is commonly used in
agriculture to genetically modify
plants, making them resistant to
insects and pests, as well as
enabling them to withstand extreme
weather conditions.
Transduction
•
Transduction is the process wherein genetically
engineered bacteriophages-viruses that parasitize
bacteria are introduced into the cell to create the
desired recombinant DNA.
Applications of Recombinant DNA
Several scientific advancements have led to many genetic engineering
techniques that are very beneficial to us. It is now possible to transfer DNA
sequences from one organism to another. American researcher Steven
Howell and his associates at the University of California in San Diego
learned that even genes from two or more different organisms can be
made to work together. Howell's team tried to isolate the gene for
luciferase an enzyme that allows fireflies to glow and insert it into tobacco
cells. When the gene was activated from the recombinant cells, the plants
glowed in the dark. This means that the basic mechanisms of gene
expression are shared by both plants and animals Here are some of the
benefits that we can get from recombinant DNA.
Agriculture
Agriculture
-Transgenic plants, which contain
genes from other organisms, play a
significant role in agriculture. Using
recombinant DNA technology, these
plants can produce natural
insecticides, reducing reliance on
synthetic pesticides. In the
Philippines, developments have led
to crops like genetically modified
maize, known as Bt corn, which
expresses a protein from the Bacillus
thuringiensis bacterium. This corn is
effective against pests like corn
borers, helping to protect crops and
benefit farmers significantly.
The following are some bacteria used in recombinant DNA technology:
1. Pseudomonas syriangae-The recombinant variant of this bacterium is
called the ice-minus bacterium, which lacks the gene responsible for ice
formation. The ice-minus bacteria prevent frost crystals from forming on
plants.
2. Pseudomonas fluorescens- This is a nonpathogenic bacterium that has the
ability to produce proteins rapidly. This characteristic is advantageous in
developing biotherapeutics and vaccines.
3. Agrobacterium tumefaciens- In its natural state, this bacterium has a
tumor-inducing (77) plasmid that causes crown gall disease in plants. The
said 7i plasmid in the bacterium can be removed and replaced with a
recombinant plasmid. This enables the now-modified bacterium to introduce
beneficial genes to plants.
Medicine
Recombinant DNA technology has significantly
impacted medicine by allowing bacteria to
produce essential substances for health and
disease treatment. For example, human insulin is
created to help diabetics, and modified human
growth hormone supports those with pituitary
gland issues, eliminating the need for risky
hormone extraction from deceased individuals.
Additionally, this technology produces important
substances like blood clotting factors for
hemophilia and hepatitis B virus surface antigens
for hepatitis B patients. Overall, recombinant DNA
technology offers many benefits in the medical
field.
Food Industry
-Recombinant technology has
enhanced the food industry by
creating crops that are resistant to
pests, diseases, and environmental
stress. This has led to increased crop
yields and reduced production costs.
Additionally, recombinant crops often
have improved nutritional quality and
longer shelf life, benefiting consumers
and producers alike.
Recombinant technology can also be
used for processing high-quality
fermented foods such as cheeses,
pickles, wines, and beers.
ESSENTIAL LEARNING
Recombinant DNA technology provides a lot of benefits
to people. So far, recombinant DNA technology has given
positive effects in agriculture and medicine. Some of its
benefits include the production of crops that are highly
resistant to drought and heat, production of vaccines to
cure various diseases, production of clotting factors and
insulin, and even gene therapy. Recombinant DNA
technology is also used for protein production, DNA
sourcing, and alternation of an organism's normal
function.
