Module 1

THE WORLD OF GENETIC ENGINEERING: What is it and

how it will change our lives in the future.
Introduction:
According to Sang Yup Lee, biotechnology is a technology that uses living organisms to make
products that allow us to implore products as diverse as household cleaning products, organs for transplant
and cleaner renewable fuels. Biotechnology is poised to change our lives, and why it could one day be as
commonplace as having a cellphone or a tablet. The genetic make-up editing of living organisms, including
microorganisms, plants and animals, is exciting for many potential applications. With these advances, we
could enhance bio-based chemicals production, increase food production and maintain a better nutritional
value, or we could manufacture organs for transplant. It is realistic to say that biotechnology will become
a part of our life, from drugs, medicine and therapeutics to environmentally friendly chemicals, fuels and
materials.

Let’s Understand
The term genetic engineering initially referred to various techniques used for the modification or
manipulation of organisms through the processes of heredity and reproduction. As such, the term
embraced both artificial selection and all the interventions of biomedical techniques, among them artificial
insemination, in vitro fertilization (e.g., “test-tube” babies), cloning, and gene manipulation. In the latter
part of the 20th century, however, the term came to refer more specifically to methods of recombinant DNA
technology (or gene cloning), in which DNA molecules from two or more sources are combined either
within cells or in vitro and are then inserted into host organisms in which they are able to propagate.

This technology raises a number of significant ethical issues. In agriculture, for instance, ethicists have
highlighted potential human health hazards associated with genetically modified crops and livestock, as
well as normative concerns about the treatment of animals and the ecological consequences of genetic
engineering. In medicine, there has been significant ethical controversy about the putative distinction
between protocols meant to restore function and those meant to enhance function beyond species-typical
norms. Additionally, ethicists have attended to the potential human health risks associated with germ-
line genetic engineering, as distinct from somatic genetic engineering. Finally, in the context of
reproduction, ethicists have argued that genetic engineering raises ethical issues involving the screening
and manipulation of embryos to eliminate or introduce various medical and/or cosmetic characteristics.

Genetic Engineering Technique

The process for genetic engineering begins the same for any organism being modified
A. Identify an organism that contains a desirable gene.
B. Extract the entire DNA from the organism.
C. Remove this gene from the rest of the DNA. One way to do this is by using a restriction enzyme. These
enzymes search for specific nucleotide sequences where they will "cut" the DNA by breaking the
bonds at this location.
D. Insert the new gene to an existing organism's DNA. This may be achieved through a number of different
Processes.
When modifying bacteria, the most common method for this final step is to add the isolated gene to
a plasmid, a circular piece of DNA used by bacteria. This is done by "cutting" the plasmid with the same
restriction enzyme that was used to remove the gene from the original DNA. The new gene can now be
inserted into this opening in the plasmid and the DNA can be bonded back together using another enzyme
called ligase. This process, shown in Figure 4, creates a recombinant plasmid. In this case, the recombinant
plasmid is also referred to as a bacterial artificial chromosome (BAC). Refer to the associated
activity Bacteria Transformation to have students create a model to simulate and learn about the process
used by genetic engineers to modify bacteria

TOOLS USED OF GENETIC ENGINEERING

A. Polymerase Chain Reaction (PCR)

Polymerase Chain Reaction (PCR) is the process of replicating multiple
copies of the genes of interest. The discovery of thermostable DNA
polymerases, such as Taq Polymerase, has made it possible to manipulate
DNA replication in the laboratory. It amplifies the quantities of DNA
segments. Primers are used to identify the gene of interest and replicate
them. These copies can then be separated and purified using gel
electrophoresis.

B. Restriction enzymes (Molecular scissor)

The discovery of enzymes known as restriction endonucleases has
been essential to protein engineering. Based on the nucleotide
sequence, these enzymes cut DNA at specific locations. DNA cut with
a restriction enzyme produces many smaller fragments of varying
sizes. These can be separated using gel electrophoresis or
chromatography.
C. Gel Electrophoresis
Purifying DNA from cell culture, or cutting it using restriction enzymes
wouldn’t be of much use if we couldn’t visualize the DNA. Gel
electrophoresis helps visualize the size and type of DNA extracted using
PCR and restriction enzymes. It is also used to detect DNA inserts and
knockouts.

D. DNA Ligase

DNA ligase can create covalent bonds between nucleotide chains. This
is done to create recombinant strands or close a circular strand that
has been cut by restriction enzymes. The enzymes DNA polymerase I
and polynucleotide kinase are also important for filling in gaps or
phosphorylating the 5′ ends, respectively.

E. Plasmid

Plasmids are small, circular pieces of DNA that

are not part of the bacterial genome but are
capable of self-replication. It is used as vectors
to transport genes between microorganisms.
Once the gene of interest has been amplified
with PCR, the gene and plasmid are cut by
restriction enzymes and ligated together. The
resulting combination is known as Recombinant
DNA. Viral (bacteriophage) DNA can also be used
as a vector, as can cosmids, which are
recombinant plasmids containing bacteriophage
genes.

F.

G. Not all cells will take up DNA during

transformation. Therefore, it is essential to
H. identify the cells that undergo a
transformation and those that have not.
I. Generally, plasmids carry genes for
antibiotic resistance, and transgenic cells
J. can be selected based on the expression of
those genes and their ability to grow on
K. media containing that antibiotic.
Alternative methods of selection depend on
L.
the presence of other reporter proteins
M. such as the x-gal/lacZ system, or green
fluorescence protein, which allow selection
N. based on color and fluorescence, res

O.

P.

Q. Identifying Transgenic Organisms