GENETIC ENGINEERING

College of Science/ biology

department
Assistant professor Dr. Munim
Radwan Ali
 Genetic Engineering
The term genetic engineering is probably the label that most people would Several terms may be
used to describe the technologies involved in manipulating genes. However, there are several other
terms that can be used to describe the technology, including gene manipulation, gene cloning,
recombinant DNA technology, genetic modification, and the new genetics. There are also legal
definitions used in administering regulatory mechanisms in countries where genetic engineering is
practiced.

 Genetic engineering is the direct manipulation of an organism's genome using biotechnology. It is a set of
technologies used to change the genetic makeup of cells, including the transfer of genes within and across species
boundaries to produce improved or novel organisms. New DNA may be inserted in the host genome by first isolating and
copying the genetic material of interest using molecular cloning methods to generate a DNA sequence, or by synthesizing
the DNA, and then inserting this construct into the host organism.
 The science of genetic engineering originated in the late 1960s, the enzyme DNA ligase was
isolated. This enzyme can join two strands of DNA together, a prerequisite for the construction of
recombinant molecules, and can be regarded as a sort of molecular glue. and early 1970s with the
discovery of restriction enzymes . The 1978 Nobel Prize for physiology went to the discoverer of
restriction enzymes, Hamilton O. Smith, and the first people to use these tools to analyze the
genetics of a virus, Daniel Nathans and Werner Arber.
 Restriction enzymes make it possible to remove a bit of DNA from one organism's chromosome
and to insert it into another organism's chromosome . This allows for the production of new
combinations of genes that may not exist in nature.
 The first recombinant DNA molecules were generated at Stanford University in 1972,
utilizing the cleavage properties of restriction enzymes (scissors) and the ability of
DNA ligase to join DNA strands together (glue).
 The importance of these first tentative experiments cannot be overestimated.
Scientists could now join different DNA molecules together and could link the DNA of
one organism to that of a completely different organism. The methodology was
tended in 1973 by joining DNA fragments to the plasmid pSC101, ensure that the
target sequence is replicated in a suitable host cell. which is an extrachromosomal
element isolated from the bacterium Escherichia coli.
 These recombinant molecules behaved as replicons; that is, they could replicate
when introduced into E. coli cells. Thus, by creating recombinant molecules in vitro,
and placing the construct in a bacterial cell where it could replicate in vivo, specific
fragments of DNA could be isolated from bacterial colonies that formed clones
(colonies formed from a single cell, in which all cells are identical) when grown on
agar plates. This development marked the emergence of the technology that
became known as gene cloning
DNA Strand cleavage by DNA
restriction enzyme fragment

Ligation the fragment with

plasmid

Cleavage the
plasmid with RE
 is any organism whose genetic material has been altered using genetic
engineering techniques (i.e. genetically engineered organism). GMOs are the source of
medicines and genetically modified foods and are also widely used in scientific research
and to produce other goods. The term more specifically defined type of GMO is a
"Transgenic Organism".
 The first GMOs were bacteria generated in 1973 and GM mice in 1974. Insulin-
producing bacteria were commercialized and the first diabetic patient in the world was
injected with human insulin made in bacteria in December 1980, making this the first
genetically engineered product to enter medical practice , and genetically modified
food has been sold since 1994. GoldFish, the first GMO designed as a pet, was first sold
in the United States in December 2003.
The interferons are another medically important group of peptides that became in abundance only after the
development of genetic engineering techniques. Interferon was useful for treating viral infections, and there were
strong indications that it might be effective against some cancers.
Before the advent of genetic engineering techniques, it took laborious processing of thousands of units of human
blood to obtain enough interferon to treat a few patients.
Other medically useful human peptides that have been made widely available because of genetic engineering are
human growth hormone, which is used to treat persons with congenital dwarfism and tissue-type plasminogen
activator (t-PA), which is a promising new treatment for persons who suffer a heart attack.
With the development of retroviral vectors in the early 1980s, the possibility of efficient gene transfer into
mammalian cells for the purpose of gene therapy became widely accepted.
 1990 deemed America to became the first country to allow new genes be
introduced into human beings . A gene drug was used to treat a 4 year-old girl
with severe combined immune deficiency (SCID). Victims of SCID the lack of
gene that controls the production commands vital to immune functioning. SCID
patients prior to gene treatment had to live inside sanitized plastic bubbles. In
early 1991, a 9 year-old girl with SCID deficiency was also treated with the same
gene therapy. In 2000 it was announced that three French infants born with SCID
had been cured using a more refined version of this technique
 A genetically modified organism (GMO) This is an organism whose genetic
makeup has been altered by the addition of genetic material from another, unrelated
organism. This should not be confused with the more general way in which "GMO" is
used to classify genetically altered organisms, as typically GMOs are organisms whose
genetic makeup has been altered without the addition of genetic material from an
unrelated organism.
 if genetic material from another species is added to the host, the resulting organism is
called transgenic. If genetic material from the same species or a species that can
naturally breed with the host is used the resulting organism is called cisgenic. Genetic
engineering can also be used to remove genetic material from the target organism,
creating a gene knockout organism
 Genetic Engineering Applications
Medicine
 Genetic engineering has resulted in a series of medical products. The first two commercially
prepared products from recombinant DNA technology were insulin and human growth hormone,
both of which were cultured in the E. coli bacteria. Since then a plethora of products have
appeared on the market, including the following abbreviated list, all made in E. coli:
 BIOnate :A vaccine is usually a harmless version of a bacterium or virus that is injected into
an organism to activate the immune system to attack and destroy similar substances in the future.
 Tumor necrosis factor. Treatment for certain tumor cells
 Interleukin-2 (IL-2). Cancer treatment, immune deficiency, and HIV infection treatment
 Prourokinase. Treatment for heart attacks
 Taxol. Treatment for ovarian cancer
 Interferon. Treatment for cancer and viral infections
 In addition, a number of vaccines are now commercially prepared from recombinant
hosts. At one time vaccines were made by denaturing the disease and then injecting it
into humans with the hope that it would activate their immune system to fight future
intrusions by that invader. Unfortunately, the patient sometimes still ended up with the
disease.
 With DNA technology, only the identifiable outside shell of the microorganism is
needed, copied, and injected into a harmless host to create the vaccine. This method is
likely to be much safer because the actual disease-causing microbe is not transferred to
the host. The immune system is activated by specific proteins on the surface of the
microorganism -e. DNA technology takes that into account and only utilizes identifying
surface features for the vaccine. Currently vaccines for the hepatitis B virus, herpes
type 2 viruses, and malaria are in development for trial use in the near future.
Agriculture :Crop plants have been and continue to be the focus of biotechnology as
efforts are made to improve yield and profitability by improving crop resistance to insects
and certain herbicides and delaying ripening (for better transport and spoilage resistance).
The creation of a transgenic plant, one that has received genes from another organism,
proved more difficult than animals. Unlike animals, finding a vector for plants proved to be
difficult until the isolation of the Ti plasmid, harvested from a tumor-inducing (Ti) bacteria
found in the soil. The plasmid is “shot” into a cell, where the plasmid readily attaches to the
plant's DNA. Although successful in fruits and vegetables, the Ti plasmid has generated
limited success in grain crops.
 Creating a crop that is resistant to a specific herbicide proved to be a success because the
herbicide eliminated weed competition from the crop plant. Researchers discovered
herbicide-resistant bacteria, isolated the genes responsible for the condition, and “shot”
them into a crop plant, which then proved to be resistant to that herbicide. Similarly, insect-
resistant plants are becoming available as researchers discover bacterial enzymes that
destroy or immobilize unwanted herbivores, and others that increase nitrogen fixation in the
soil for use by plants.
 Geneticists are on the threshold of a major agricultural breakthrough. All plants need
nitrogen to grow. In fact, nitrogen is one of the three most important nutrients a plant
requires. Although the atmosphere is approximately 78 percent nitrogen, it is in a form
that is unusable to plants. However, a naturally occurring rhizobium bacterium is found
in the soil and converts atmospheric nitrogen into a form usable by plants. These
nitrogen-fixing bacteria are also found naturally occurring in the legumes of certain
plants such as soybeans and peanuts. Because they contain these unusual bacteria, they
can grow in nitrogen-deficient soil that prohibits the growth of other crop plants.
Researchers hope that by isolating these bacteria, they can identify the DNA segment
that codes for nitrogen fixation, remove the segment, and insert it into the DNA of a
profitable cash crop! In so doing, the new transgenic crop plants could live in new fringe
territories, which are areas normally not suitable for their growth, and grow in current
locations without the addition of costly fertilizers!
 Projects with genetically modified products
 DHA canola: We have developed canola plants which produce high quality oils
rich in omega-3 DHA (docosahexaenoic acid). This nutrient is currently only found
in beneficial quantities in ocean-based algae, and the fish that eat it. This product
could break the world's reliance on fish stocks while meeting the increasing
demand for these healthy long-chain omega-3 oils.
 Leaf oil: We have engineered tobacco plants to have oilseed-like levels of oil in
their leaves (around 35 per cent). This product could provide an economically
competitive renewable alternative to petroleum diesel.
 BT cowpeas: We are part of a global project to improve cowpea production in
Africa and are making progress towards incorporating ‘built-in’ insect pest
protection that could help reduce food shortages in some African regions.
 Animal Husbandry
 Neither the use of animal vaccines nor adding bovine growth hormones to
cows to dramatically increase milk production can match the real excitement in
animal husbandry: transgenic animals and clones.
 Transgenic animals model advancements in DNA technology in their
development. The mechanism for creating one can be described in three steps:
 Healthy egg cells are removed from a female of the host animal and
fertilized in the laboratory.
 The desired gene from another species is identified, isolated, and cloned
 The cloned genes are injected directly into the eggs, which are then
surgically implanted in the host female, where the embryo undergoes a
normal development process.
 It is hoped that this process will provide a cheap
and rapid means of generating desired enzymes,
other proteins, and increased production of meat,
wool, and other animal products through common,
natural functions.
 Ever since 1997 when Dolly was cloned, research
and experimentation to clone useful livestock has
continued unceasingly. The attractiveness of cloning
is the knowledge that the offspring will be genetically
identical to the parent as in asexual reproduction.
Four steps describe the general process:
 A differentiated cell, one that has become specialized during
development, with its diploid nucleus is removed from an animal to
provide the DNA source for the clone.
 An egg cell from a similar animal is recovered and the nucleus is
removed, leaving only the cytoplasm and cytoplasm organelles.
 The two egg cells are fused with an electric current to form a single
diploid cell, which then begins normal cell division.
 The developing embryo is placed in a surrogate mother, who then
undergoes a normal pregnancy.