Genetics,

Variation and
Natural Selection
Genetic Engineering

Done by: Dequan Holder

Restriction Enzymes and DNA Ligase

• Restriction enzymes are DNA-cutting enzymes. Each enzyme recognizes

one or a few target sequences and cuts DNA at or near those sequences.
• Many restriction enzymes make staggered cuts, producing ends with
single-stranded DNA overhangs. However, some produce blunt ends.
• DNA ligase is a DNA-joining enzyme. If two pieces of DNA have matching
ends, ligase can link them to form a single, unbroken molecule of DNA.
• In DNA cloning, restriction enzymes and DNA ligase are used to insert
genes and other pieces of DNA into plasmids.
• Restriction enzymes and DNA ligase are often used to insert genes and
other pieces of DNA into plasmids during DNA cloning.
Diagram depicting restriction process.
Diagram depicting the DNA ligase process.
Steps in recombinant DNA (rDNA) technology.

• Recombinant DNA (rDNA) technology refers to the process of joining

DNA molecules from two different sources and inserting them into a
host organism, to generate products for human use. This process
consist of a number of steps which are:
• Isolation of genes
• Restriction enzyme digestion
• Cloning of genes
• Ligation of DNA
• Insertion of recombinant Dna into the host
Isolation of Genes

• Gene isolation, which is the isolation of a particular gene from the

rest for use in genetic engineering.
• The cell containing the gene is opened and the DNA is purified.The
gene is separated by using restriction enzymes to cut the DNA into
fragments or polymerase chain reaction (PCR) to amplify up the
gene segment.
Restriction Enzyme Digestion

• Restriction enzymes act as molecular scissors that cut DNA at

specific locations.
• They involve the incubation of the purified DNA with the selected
restriction enzyme, at conditions optimal for that specific enzyme.
Ligation of DNA Molecules

• The purified DNA and the vector of interest are cut with the same
restriction enzyme.
• This gives us the cut fragment of DNA and the cut vector, that is
now open.
• The process of joining these two pieces together using the enzyme
‘DNA ligase’ is ‘ligation’. The resulting DNA is ‘recombinant DNA‘.
Insertion of Recombinant DNA Into Host

• The recombinant DNA is introduced into a recipient host cell. This

process is ‘Transformation’.
• Bacterial cells do not accept foreign DNA easily. Therefore, they
are treated to make them ‘competent’ to accept new DNA
An Application of Genetic Engineering

Insulin Production
Bacterial cells can be genetically modified so that they have the
gene for producing human insulin. As these modified bacteria grow,
they produce human insulin. This protein can be purified and
supplied to diabetics.
Gene Therapy
Gene therapy is an experimental technique that uses genes to treat or
prevent disease. There are three (3) known approaches to gene therapy
which are:
• Replacing a mutated gene that causes disease with a healthy copy of the
gene.
• Inactivating, or “knocking out,” a mutated gene that is functioning
improperly.
• Introducing a new gene into the body to help fight a disease.
Although gene therapy is a promising treatment option for a number of
diseases (including inherited disorders, some types of cancer, and certain
viral infections), the technique remains risky and is still under study.
Application in Gene Therapy.

• Cystic fibrosis results from a mutation in the carrier protein (CFTR)

which pumps chloride ions out of cells in the respiratory tract and
alimentary canal.
• Person who are homozygous recessive for the disorder have thick
and sticky mucus.
• Liposomes and viruses have been used as vectors to deliver the
dominant allele into the cells of respiratory treact using an inhaler.
• This had mixed success as the vectors tend to stimulate
immflammation and some clinical trials had to be stopped.
What are GMOs?
• GMO stands for “Genetically Modified Organism,” and the phrase refers
to plants or animals that have been genetically engineered to produce a
specific set of characteristics.
• DNA is taken from a completely different species (like a virus or fish) and
inserted into a crop (like a carrot) to transfer genetic traits. These
experimental combinations of genes do not occur in nature.
• The use of GMOs in the environment also comes with implications which
can be medical, agricultural, ethical and social.
Implications involving the usage of GMOs in
the Environment.

Agriculturally
• Out-crossing is the unintentional breeding of a domestic crop with a
related plant.
• A major environmental concern associated with GM crops is their
potential to create new weeds through out-crossing with wild relatives.
• Managing weeds is one of the most tedious tasks of farming.
• scientists developed genetically modified herbicide-tolerant (HT) crops
so farmers can spray their fields with weed killers without affecting their
crop yield.This led to the use of glyphosate herbicides. Overtime the
weeds developed aresistance to the herbicide.  
Implications involving the usage of GMOs in
the Environment cont’d

Ethically
• Antibiotic resistance factors used to identify GMOs could escape
and be transferred to pathogenic organisms.
• Herbicide resistance genes could be transferred from pollen to
weed species and lead to the development of superweeds that are
resistant to herbicides.
• Foreign genes can pollute non-GM and organic crops which require
certification that they provide GM-free food.
Implications involving the usage of GMOs in
the Environment cont’d

Medically
• For many years diabetes was treated by regular injections of
insulin which was extracted from animals.but now it is produced
by GM bacteria or yeasts.
• The main disadvantage of GM human insulin is that some people
• Have reported that they do not experience any warning signs of a
hypoglycaemic attack when the blood glucose concentration falls
increasing the likelihood of a diabetic coma.
END OF PRESENTATION!!!

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