BASIC SCIENTIFIC

OF BIOTECHNOLOGY

Dr. Rahem Al.Ziadi

What Is Biotechnology and What Does It
Mean to You?
• Biotechnology – using living organisms,
or the products of living organisms, for
human benefit to make a product or solve
a problem

• Historical Examples
– Fermentation
– Selective breeding
– Use of antibiotics
 • Example of Biotechnology – Selective Breeding

(a) (b)

Normal zebrafish "Casper" zebrafish – made

by selective breeding
• What feature of Casper makes it a "model organism" to study migration of
cancer cells compared to wildtype fish?

Via selective breeding, ‫ التساوج االنتقالي‬by mating a zebrafish mutant that lacked reflective pigment
with a zebrafish that lacked black pigment, Casper is transparent. This feature allows scientists to
inject fluorescent cancer cells into Casper's abdominal cavity and then track the migration of
those cells to specific locations in his body.
• Based on this tree,
can you become
successful in the
biotech industry
only studying
biology?

No, you need to take many science

courses including biology,
computer science, chemistry, math
(especially statistics). If you are not
broadly trained, you will not be
able to obtain a job in this growing
field!
• Modern Examples
– Gene cloning
– Genetic engineering
– Recombinant DNA technology
– Human Genome Project
• Example of "modern" biotechnology:
– recombinant DNA technology started modern biotech
as an industry
• Examples of applications
– development of disease-resistant plants
– food crops that produce greater yields
– "golden rice" engineered to be more nutritious
– genetically engineered bacteria that can degrade
environmental pollutants
• Gave more examples of applications‫؟‬

:.
-Low cost production of proteins used to treat diseases
-Development of recombinant enzymes to be used by molecular biologists
- for research
• Look at the two chromosomes and determine which chromosome has
more than one gene involved in promoting breast cancer. Explain your
answer.

– Now use the link to further study the diseases involved in these chromosomes
• http://ghr.nlm.nih.gov/chromosome

chromosome 13 has 2 genes listed regarding breast cancer and chromosome 21 only has 1 gene
related to breast cancer.
 • Most drugs are developed to combat ‫ محاربة‬diseases affecting humans
– Why?
• Which disease has the most drug candidates ‫ ? مرشحٌن‬Why does that
disease have more drug candidates than hepatitis C?

1. This reflects the current needs of humans- we have too many diseases and currently too few drugs to target them.

2. By far, cancer has the most drug candidates than any other disease. This disease has many more drug candidates
than hepatitis C because hepatitis C affects less people worldwide than different kinds of cancer. Cancer, of course,
can affect many different organs. On the other hand, hepatitis C only affects the liver.
• Use genetically modified cultured cells to
make protein ‫؟‬
• Products of Modern Biotechnology
– Example of proteins created by gene cloning
called recombinant proteins
 Types of Biotechnology

• Microbial Biotechnology
• Agricultural Biotechnology
• Animal Biotechnology
• Forensic Biotechnology
• Bioremediation
• Aquatic Biotechnology
• Medical Biotechnology
• Regulatory Biotechnology
Types of Biotechnology

• Microbial Biotechnology –
manipulation ‫ معالجة‬of microorganisms such
as yeast and bacteria
– Create better enzymes
– More efficient decontamination processes for
industrial waste product removal
– Used to clone and produce large amounts of
important proteins used in human medicine
 Types of Biotechnology

• Agricultural Biotechnology
– United Nations Food and Agricultural Org.
predicts by 2050, we will need to feed a world
population of 9.1 billion! This requires raising
food production by approximately 70%!
– brainstorm : gave a few solutions to better feed the
world by 2050?

1. use of recombinant technology to genetically modify foods. For example, scientists can
create ‫ ايجاد‬better crops that are perhaps more drought ‫ جفاف‬resistant.
2. They can create ways ‫ خلك طرق‬to better preserve ‫ حفظ‬fruits and vegetables so they don't
quickly rot ‫ تتعفن‬and get wasted ‫ تتلف‬.
3. Perhaps create crops that are foul tasting ‫ طعم غير مستساغ‬to pests ‫ االفات‬including bugs and
animals so that the crops survive.
4. Perhaps create ways to have crops grow even in the most extreme conditions.
5. obtain proteins from other novel food sources.
Types of Biotechnology

• Agricultural Biotechnology
– Plants more environmentally friendly that yield
more per acre ‫( دونم‬genetically engineered)
– Resistance to diseases and insects
– Foods with higher protein or vitamin content
– Drugs developed and grown as plant products
– These better plants ultimately reduce
production costs to help feed the growing
world population
 Types of Biotechnology

• Agricultural Biotechnology

– discuss how you can use this technology

in a third world country to create a better
corn crop (main crop in that country) that
contains all of the 22 essential amino
acids‫؟‬
 Types of Biotechnology

• Will improved crops ‫ المحاصٌل المحسنة‬that are

created to satisfy ‫ لتلبٌة‬world hunger
‫الجوع‬reduce available ‫ متاحة‬land for
biofuel‫ وقود حٌوي‬crops? Discuss

This is an ethical question and should lead to some powerful

discussions.
discuss what is meant by biofuel crops
 Types of Biotechnology

• Animal Biotechnology
– Animals as a source of medically valuable
proteins
• Antibodies
• Transgenic animals ‫معدلة وراثٌا‬
– Animals as important models in basic
research
• Gene "knockout" experiments ‫تضرٌب‬
• Design and testing of drugs and genetic therapies
– Animal cloning
• Source of transplant organs ‫زراعة أعضاء‬
 Types of Biotechnology

• Animal Biotechnology
– transgenic animal: way to achieve‫ لتحقٌق‬large
scale production of therapeutic proteins from
animals for use in humans
– Female transgenic animals express
therapeutic proteins in milk (contains genes
from another source)
– Example: human genes coding for clotting‫تخثر‬
proteins can be introduced into female goats
for production of these proteins in their milk
 Types of Biotechnology
• Animal Biotechnology
– Gene knockout: (gene knock-in)
• Disrupt ‫ تعطٌل‬a gene in the animal and then look at what
functions are affected in the animal as a result of the loss of
the gene
• This allows ‫ ٌسمح‬researchers to determine the role and
function of the gene
• Since humans are similar to rats and mice, gene knockout
studies in rats and mice can lead to better understanding of
gene function in humans.
• give an example of a gene you would like to knockout in mice?

Perhaps, you want to study the role of insulin in preventing ‫منع‬diabetes. By knocking out
the gene coding for insulin, you can observe ‫ رصذ‬the animal's response (development of
diabetes). Then scientists can inject recombinant insulin to the animal and again, study
their responses to the treatment
• Forensic Biotechnology)ً‫تقنٌات الطب الشرعً (الجنائ‬
– DNA fingerprinting
• Inclusion or exclusion of a person from suspicion ‫الشك‬
Paternity cases ‫االبوة‬
• Identification of human remains ‫تحدٌد الرفاة‬
• Endangered species‫االنواع المهددة باالنقراض‬
• Tracking and confirmation of the spread of disease ‫تتبع االمراض‬
Types of Biotechnology

• Forensic Biotechnology
• Based on DNA results
from this gel, did the
defendant ‫ المدعى علٌه‬commit ‫ارتكب‬
this crime ‫ ?جرٌمة‬Explain based
on the gel results.

Based on the gel results, there is evidence that

the defendant committed the crime. The
position of the bands as well as number of
bands on the gel match with the victim's
‫ضحيت‬blood. It is important to note, that though
the bands are very light for the sample from the
jeans, they also match the victim's blood.
he figure below represents the results of a DNA gel electrophoresis of DNA collected at a crime scene, a sample
from the victim, and samples from suspects. Use the figure to determine which, if any, suspects' DNA matches the
crime scene sample. Explain your reasoning.
Example : Compare the DNA collected from the crime scene to determine which of the three
suspects was present (click picture to solve)
Example: Paternal Testing
Compare the profiles of three men with that of a mother and child to determine the biological
father (click picture to solve)
Example: Paternal Testing
 Types of Biotechnology

• Bioremediation‫الكنس الحٌوي‬
– The use of biotechnology to process and
degrade a variety of natural and man-made
substances ‫المواد االصطناعٌة‬
• Particularly those that contribute ‫تساهم‬to
environmental pollutionً‫التلوث البٌئ‬
– Example – stimulated ‫ تحفٌز‬growth of bacteria
that degrade components in crude oil
• 1989 Exxon Valdez oil spill ‫ تسرب‬in Alaska
• 2010 Deep Water Horizon spill promoted research
into natural oil-degrading organisms and enzymes
Types of Biotechnology

• Bioremediation – adding nutrients to stimulate

growth of bacteria to clean up oil spill
Q\\ Describe how bacteria are used in pollution reduction
operations (oil-eating bacteria) in the oceans?
 Types of Biotechnology
• Aquatic Biotechnology
• Why create ‫ يخلق‬transgenic salmon overproducing growth
hormone?
transgenic

normal

Two different salmon

• How does this modified salmon help humans?
Answer 1.
By creating transgenic salmon that overproduce Growth Hormone, it allows the salmon to have
fast growth rates over a short period of time.
Answer 2.
This modified salmon helps humans because it decreases the time and expenses required to grow
the salmon for market sale.
 Types of Biotechnology

• Medical Biotechnology
1. Involved with the whole spectrum ‫كل‬
‫طٌف‬of human medicine
• Preventive medicine ‫ادوٌة وقائٌة‬
• Diagnosis of health and illness‫تشخٌص الصحة والمرض‬
• Treatment of human diseases ‫عالج االمراض البشرٌة‬
2. New information from Human Genome
Project
• Gene therapyً‫العالج الجٌن‬
3. Stem cell ‫ الخالٌا الجذعٌة‬technologies
• Stem cells – grown in lab
and then treated with
different chemicals to
allow ‫ يسمح‬them to develop
into specific kinds of
tissues needed for
transplant ‫زراعت األعضاء‬
• Current use ‫االستعمال الحالي‬:
stem cells are used for
diabetes; spinal cord
injuries ‫اصاباث الحبل الشوكي‬
• Can you gave other diseases that scientists are testing with stem
cells?
• replace neurons damaged by spinal cord injury, stroke, Alzheimer’s disease, Parkinson’s disease or other
neurological problems;
• produce insulin that could treat people with diabetes and heart muscle cells that could repair damage after a
heart attack; or
• replace virtually any tissue or organ that is injured or diseased
Remember, most of these diseases are only be tested with stem cell therapy in animal models. They
are not being used in the clinic ‫العياداث‬yet!!!
Q\\explain why scientists are doing more research using
embryonic vs. adult stem cells.
• Medical biotechnology
– Genes are headline ً‫عنوان رئٌس‬news items
• How will medical biotechnology change our lives in the
years ahead ‫? قادمة‬
– Human Genome Project ‫مشروع الجٌنوم البشري‬
• Research on the function of human genes and controlling factors that
regulate genes
– Human proteome ‫بروتٌنوم البشر‬
• Group of proteins responsible for activity in a
human cell
• How will medical biotechnology change our lives in the years ahead?
– Single Nucleotide Polymorphisms (SNPs)
• Single nucleotide changes ‫(تتغٌر‬mutations) in DNA sequences that vary ‫ تختلف‬from
individual ‫ فرد‬to individual
(SNP :is a variation in a single nucleotide that occurs at a specific position in the
genome)
• These variations ‫االختالفات‬are the cause ‫ سبب‬of some genetic diseases ‫االمراض الوراثٌة‬
(sickle cell anemiaً‫)فقر الدم المنجل‬
• SNPs will help identify genes involved ‫ مشاركة‬in medical conditions ً‫حاالت مرض‬
including arthritis ‫التهاب المفاصل‬, stroke ‫سكته دماغٌة‬, cancer, heart disease, diabetes, and
behavioral ‫ السلوكٌة‬and emotional ‫ النفسٌة‬illnesses
• Example of SNPs and breast cancer
• Identification of SNPs in BRCA1 and BRCA2 genes
involved ‫ مشاركت‬in promoting‫ تعسيس‬breast cancer led ‫ قاد‬to
development‫ تطوير‬of better targeted treatments for
people who have those specific gene mutations ‫طفراث‬
‫جينيت محذدة‬
• Can you think of how this knowledge might be useful for someone who
is not already diagnosed with cancer? (think of your basic knowledge
of genetics.)

Assume‫ افترض‬that friends mom and sister both had breast cancer and found out that
they both had the BRCA1 gene mutation. His doctor would counsel he to be tested for
this mutation too. Then, if he has the mutation, the doctor could monitor he more
closely for developing breast cancer. If he was to develop this cancer, they would be
able to catch it early on when he has a better chance of being treated successfully. It is
important to note that even if he has tested and have this genetic mutation, it does NOT
mean that he will definitely develop cancer. It just means that he has an increased risk!
It is also important to note that both men and women can develop breast cancer
 • Example of how we can benefit ‫نستفاد‬from the human genome project

• Based on the figure ‫اعتمادا على الشكل‬, why doesn't person 2 develop a genetic
disease due to the SNP (G → T)?
Since there can be multiple codons coding for the same amino acid, then even if person 2 has a SNP, the new
codon can still code for the same amino acid as in person 1. Thus, the same protein will ultimately be made
and it will therefore have the correct function. This type of mutation is called silent since the nucleotide
change did not affect a change in the amino acid sequence.
Person 3 had a SNP that changed the codon so it coded for a different amino acid. Thus, this will affect the
way the protein is folded ‫ التفاف‬and, if not in the proper structure ‫التركيب الصحيح‬, it will have impaired ‫ضعف‬
function or perhaps no function.
• Gene therapy technology
– Replacing ‫استبدال‬or augmenting ‫ اضافة‬defective genes ‫ جٌن معٌن‬with
normal copies of the gene

Still have barriers ‫ تحدٌات‬to overcome before this technology becomes safe and
effective
• Obstacles include ‫تشمل العقبات‬:
– How can normal genes be delivered to all cells in the body?
– What are the long-term effects ‫االثار على المدى الطوٌل‬of introducing ‫ ادخال‬extra ‫جٌنات‬
‫ إضافٌة‬genes in humans?
– What must be done to ensure‫ ضمان‬the proper protein is made after the genes
are delivered to the body?
• How will medical biotechnology change
our lives in the years ahead?
– Regenerative medicine ‫الطب التجدٌدي‬
• Genetically modifying stem cells of patients to
treat genetic disease
• In future scientists will be able to…
1. Isolate adult stem cells from a patient with a
genetic disorder ‫اضطرابات وراثٌة‬
2. Genetically manipulate ‫ تالعب‬these cells by gene
therapy
3. Reinsert ‫عادة ادخال‬the cells into the same patient to
help and treat their genetic disease
Insulin
Insulin is one of the important pharmaceutical products produced commercially by genetically
engineered bacteria. Before this development, commercial insulin was isolated from animal
pancreatic tissue. Microbial insulin has been available since 1982. The human
insulin gene is introduced into a bacterium like E. coli. Two of the major advantages of insulin
production by microorganisms are that the resultant insulin is chemically identical
to human insulin, and it can be produced in unlimited quantities.
What is genetic engineering and what is it
used for?
The ability to manipulate and analyse DNA using genetic engineering
techniques (recombinant DNA technology) was foreseen in the
mid 1960s and came to fruition in the early 1970s. The technology,

The basic tools of genetic engineering

The techniques for isolating, cutting and joining molecules of DNA,
developed in the early 1970s, have provided the foundations of our
current technology for engineering and analysing nucleic acids. These
allow fragments of DNA from virtually any organism to be cloned in
a bacterium by inserting them into a vector (carrying) molecule that
is stably maintained in the bacterial host.
Isolation and purification of nucleic acids
Biochemical techniques for preparing large quantities of relatively
pure nucleic acids from microbial cells are an essential pre-requisite
for in-vitro gene technology. The first step in the isolation of nucleic
acids is the mechanical or enzymatic disruption of the cell to release
the intracellular components that include the nucleic acids. Once
released from the cell, the nucleic acids must be purified from other
cellular components such as proteins and polysaccharides to provide
a substrate of appropriate purity for nucleic acid modifying enzymes.
The released nucleic acids are recovered using a combination of
techniques including centrifugation, electrophoresis, adsorption to inert
insoluble substrates or precipitation with non-aqueous solvents
Cutting DNA molecules
The ability to cut molecules of DNA, either randomly or at specific
target sites, is a requirement for many recombinant DNA techniques.
DNA may be cleaved using mechanical or enzymatic methods.
Mechanical shearing is non-specific and results in the production of
random DNA fragments, which are often used to generate genomic
libraries (see Section 4.4.5). When DNA molecules are mechanically
sheared it is not possible to isolate a specific fragment containing,
for example, a particular gene or operon. In contrast, when the DNA
is cut using restriction endonucleases, which recognize and cleave
specific target base sequences in double-stranded (ds)DNA, specific
fragments can be isolated. Restriction endonucleases cut the
phosphodiester backbone of both strands of the DNA to generate 3OH
and 5PO4 termini. Several hundred restriction endonucleases have
been isolated from a wide variety of microbial species. Restriction
endonucleases are classified into groups with distinct biochemical
properties; type II enzymes are the main class used for genetic
engineering purposes
Restriction endonucleases are named according to the species from which they were originally
isolated; enzymes isolated from Haemophilus influenzae are designated Hin, those from Bacillus
amyloliquefaciens, Bam, etc. When more than one type of enzyme is isolated from a particular
strain or species, the strain and isolation number (in roman numerals) are added to the name.
Thus the three restriction endonucleases isolated from H. influenzae strain Rd are designated
HindI, HindII and HindIII.