Banquerohan legazpi city

Banquerohan national high school (BNHS)

PROJECT
IN
BIOTECH
Submitted to:
Mrs. alma zaragosa

Submitted by:
CHRISTINE ANN NODALO
ella mae onato

Genetic engineering
Genetic engineering, also called genetic modification, 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 cloningmethods to generate a DNA
sequence, or by synthesizing the DNA, and then inserting this construct into the host
organism. Genes may be removed, or "knocked out", using a nuclease. Gene targeting is a different
technique that uses homologous recombination to change an endogenous gene, and can be used to
delete a gene, remove exons, add a gene, or introduce point mutations.
An organism that is generated through genetic engineering is considered to be
a genetically modified organism (GMO). The first GMOs were bacteria generated in 1973 and GM
mice in 1974. Insulin-producing bacteria were commercialized in 1982 and genetically modified
food has been sold since 1994. GloFish, the first GMO designed as a pet, was first sold in the United
States in December 2003.

Genetic engineering techniques have been applied in numerous fields including research, agriculture,
industrial biotechnology, and medicine. Enzymes used in laundry detergent and medicines such as
insulin and human growth hormone are now manufactured in GM cells, experimental GM cell lines
and GM animals such as mice or zebrafish are being used for research purposes, and genetically
modified crops have been commercialized.

Definition
Genetic engineering alters the genetic make-up of an organism using techniques that
remove heritable material or that introduce DNA prepared outside the organism either directly into the host or into a cell
that is then fused or hybridized with the hostThis involves usingrecombinant nucleic acid (DNA or RNA) techniques to
form new combinations of heritable genetic material followed by the incorporation of that material either indirectly through
a vector system or directly through micro-injection, macro-injection and micro-encapsulationtechniques.
Genetic engineering does not normally include traditional animal andplant breeding, in vitro fertilisation, induction
of polyploidy, mutagenesisand cell fusion techniques that do not use recombinant nucleic acids or a genetically modified
organism in the process.[ However the European Commission has also defined genetic engineering broadly as
including selective breeding and other means of artificial selection.Cloning and stem cell research, although not
considered genetic engineering,are closely related and genetic engineering can be used within them. [7] Synthetic
biology is an emerging discipline that takes genetic engineering a step further by introducing artificially synthesized
material from raw materials into an 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 knockoutorganism. In Europe genetic modification is synonymous with genetic
engineering while within the United States of America it can also refer to conventional breeding methods. The Canadian
regulatory system is based on whether a product has novel features regardless of method of origin. In other words, a
product is regulated as genetically modified if it carries some trait not previously found in the species whether it was
generated using traditional breeding methods (e.g.,selective breeding, cell fusion, mutation breeding) or genetic
engineering.Within the scientific community, the termgenetic engineering is not commonly used; more specific terms such
as transgenic are preferred.

Genetically modified
organisms
Plants, animals or micro organisms that have changed through genetic engineering are termed genetically
modified organisms or GMOs.Bacteria were the first organisms to be genetically modified. Plasmid DNA containing new
genes can be inserted into the bacterial cell and the bacteria will then express those genes. These genes can code for
medicines or enzymes that process food and other substrates.Plants have been modified for insect protection, herbicide
resistance, virus resistance, enhanced nutrition, tolerance to environmental pressures and the production of edible
vaccines. Most commercialised GMO's are insect resistant and/or herbicide tolerant crop plants. Genetically modified
animals have been used for research, model animals and the production of agricultural or pharmaceutical products. They
include animals with genes knocked out, increased susceptibility to disease, hormones for extra growth and the ability to
express proteins in their milk.

History
Humans have altered the genomes of species for thousands of years through selective breeding, or artificial selection1 as
contrasted with natural selection, and more recently through mutagenesis. Genetic engineering as the direct manipulation
of DNA by humans outside breeding and mutations has only existed since the 1970s. The term "genetic engineering" was
first coined by Jack Williamson in his science fiction novel Dragon's Island, published in 1951[ one year before DNA's
role in heredity was confirmed by Alfred Hershey and Martha Chase, and two years before James Watson and Francis
Crick showed that the DNA molecule has a double-helix structure though the general concept of direct genetic
manipulation was explored in rudimentary form in Stanley G. Weinbaum's 1936 science fiction story Proteus Island.

Process
The first step is to choose and isolate the gene that will be inserted into the genetically modified organism.
As of 2012, most commercialised GM plants have genes transferred into them that provide protection against insects or
tolerance to herbicides. The gene can be isolated using restriction enzymes to cut DNA into fragments and gel
electrophoresis to separate them out according to length. Polymerase chain reaction (PCR) can also be used to amplify
up a gene segment, which can then be isolated through gel electrophoresis. If the chosen gene or the donor
organism's genome has been well studied it may be present in a genetic library. If the DNA sequence is known, but no
copies of the gene are available, it can be artificially synthesized.The gene to be inserted into the genetically modified
organism must be combined with other genetic elements in order for it to work properly. The gene can also be modified at
this stage for better expression or effectiveness. As well as the gene to be inserted most constructs contain
a promoter and terminator region as well as a selectable marker gene. The promoter region initiates transcription of the
gene and can be used to control the location and level of gene expression, while the terminator region ends transcription.
The selectable marker, which in most cases confers antibiotic resistance to the organism it is expressed in, is needed to
determine which cells are transformed with the new gene. The constructs are made using recombinant DNA techniques,
such as restriction digests, ligations and molecular cloning.The manipulation of the DNA generally occurs within
a plasmid.

The most common form of genetic engineering involves inserting new genetic material randomly within the host genome.
[
Other techniques allow new genetic material to be inserted at a specific location in the host genome or generate
mutations at desired genomic loci capable of knocking out endogenous genes. The technique of gene
targetinguses homologous recombination to target desired changes to a specific endogenous gene. This tends to occur
at a relatively low frequency in plants and animals and generally requires the use of selectable markers. The frequency of
gene targeting can be greatly enhanced with the use of engineered nucleases such as zinc finger
nucleases,engineeredhoming endonucleases,or nucleases created from TAL effectors.In addition to enhancing gene
targeting, engineered nucleases can also be used to introduce mutations at endogenous genes that generate a gene
knockout.

Transformation

A. tumefaciens attaching itself to a carrot cell

Only about 1% of bacteria are naturally capable of taking up foreign DNA. However, this ability can be induced in other
bacteria via stress (e.g. thermal or electric shock), thereby increasing the cell membrane's permeability to DNA; up-taken
DNA can either integrate with the genome or exist as extrachromosomal DNA. DNA is generally inserted into animal cells
using microinjection, where it can be injected through the cell's nuclear envelopedirectly into the nucleus or through the
use of viral vectors.] In plants the DNA is generally inserted using Agrobacterium-mediated
recombination or biolistics.In Agrobacterium-mediated recombination, the plasmid construct contains T-DNA, DNA which
is responsible for insertion of the DNA into the host plants genome. This plasmid is transformed
into Agrobacterium containing no plasmids prior to infecting the plant cells. The Agrobacterium will then naturally insert
the genetic material into the plant cells. In biolistics transformation particles of gold or tungsten are coated with DNA and
then shot into young plant cells or plant embryos. Some genetic material will enter the cells and transform them. This
method can be used on plants that are not susceptible toAgrobacterium infection and also allows transformation of plant
plastids. Another transformation method for plant and animal cells is electroporation. Electroporation involves subjecting
the plant or animal cell to an electric shock, which can make the cell membrane permeable to plasmid DNA. In some
cases the electroporated cells will incorporate the DNA into their genome. Due to the damage caused to the cells and
DNA the transformation efficiency of biolistics and electroporation is lower than agrobacterial mediated transformation
and microinjection.As often only a single cell is transformed with genetic material the organism must be regenerated from
that single cell. As bacteria consist of a single cell and reproduce clonally regeneration is not necessary. In plants this is
accomplished through the use of tissue culture. Each plant species has different requirements for successful
regeneration through tissue culture. If successful an adult plant is produced that contains the transgene in every cell. In
animals it is necessary to ensure that the inserted DNA is present in the embryonic stem cells. Selectable markers are
used to easily differentiate transformed from untransformed cells. These markers are usually present in the transgenic
organism, although a number of strategies have been developed that can remove the selectable marker from the mature
transgenic plant.When the offspring is produced they can be screened for the presence of the gene. All offspring from the
first generation will be heterozygous for the inserted gene and must be mated together to produce a homozygous animal.
Further testing uses PCR, Southern hybridization, and DNA sequencing is conducted to confirm that an organism
contains the new gene. These tests can also confirm the chromosomal location and copy number of the inserted gene.
The presence of the gene does not guarantee it will be expressed at appropriate levels in the target tissue so methods
that look for and measure the gene products (RNA and protein) are also used. These include northern hybridization,
quantitative RT-PCR,Western blot, immunofluorescence, ELISA and phenotypic analysis. For stable transformation the
gene should be passed to the offspring in a Mendelian inheritance pattern, so the organism's offspring are also studied.

Genetic engineering and proteomics

enetic engineering is any process in which an organisms genome is intentionally altered.

Genetic engineering does not encompass traditional breeding techniques because it requires
manipulation of an organisms genes
through cloning or transformation via the addition of foreign DNA. This

Process has five steps:

1. Isolation of the genes
2. Insertion of those genes into a transfer vector (a virus or a plasmid used
as a conduit)
3. Transfer of the vector to the organism to be modified
4. Transformation of that organisms cells
5. Separation of the genetically modified organism (GMO) from
organisms that have not been successfully modified
Proteomics is the study of proteins, and it represents the other side of
genomics. Proteins are the substance within cells that allow for the cells
various functions. They are organic compounds comprised of a chain of
molecules called amino acids that are joined by a chemical bond. While
an organisms genome remains constant across the bodys many different types of cells, its proteome,
the particular proteins expressed by the
genome, shifts from cell to cell, making proteomics more complicated than
genomics.
The field of proteomics is headed by an international consortium of
scientists and organizations known as the human Proteome Organization,
which was formed in 2001 to capitalize on the discoveries of the human
Genome Project. The science of pharmocogenomics, which combines the
fields of genomics and proteomics, may lead to custom-made treatments for
individuals based on their genetic analysis. Scientists hope that pharmocogenomics will eliminate much of the
trial-and-error process patients currently
undergo to find optimal medication dosages.

BIOTEChNOlOGy AND GENETIC

ENGINEERING
The story of insulin
Diabetes mellitus is a disease in which the body cannot produce enough insulin to
regulate blood sugar, or glucose, levels. Blood sugar levels that are either too high or too low can result in
significant health problems, including loss of consciousness and brain, kidney, eye, and nerve damage.
Synthetically produced insulin was one of the earliest success stories of molecular medicine. Type 2
diabetes, the most common variety, is usually caused by obesity, a high-fat diet, and a lack of physical
activity. Symptoms can include an impaired cardiovascular system, blindness, and renal failure. The
CDChas declared Type 2 diabetes to be an epidemic in the United States, and the disease is being
diagnosed more frequently in young people than it used to be. Individuals may also inherit a genetic
predisposition toward Type 2 diabetes.The treatment for diabetes includes insulin shots. Insulin is a hormone
normally produced by the pancreas that allows the bodys cells to obtain glucose from blood cells and store it
as glycogen in the liver and muscles. Without insulin, a person will die. historically, diabetics received shots of
insulin derived from cows, horses, pigs, or fish. But in 1982 the first synthetic insulin, humulin, came on the
market; it was genetically engineered by Genentech and marketed by Eli lilly. humulin was the first
medication genetically engineered by inserting human DNA into a host cell, which then reproduced with the
correct form of insulin to be injected into the diabetic. This synthetic insulin has proven invaluable as rates of
diabetes skyrocket.genetic screening: eliminating tay-sachs disease GM crops have become popular in recent
years for three main reasons. First and foremost, they have higher yields than nonGM crops. For example,
prior to 1950, farmers typically harvested 40 bushels of corn per acre, whereas GM corn produces up to
180 bushels per acre 4
This has important ramifications in a world that is increasingly urban, where fewer farmers supply food for a
growing population. Second, GM seed is designed to resist pests and herbicides. GM corn crops, such as

Monsantos Roundup Ready Bt corn, can be sprayed with Monsantos Roundup herbicide and continue to grow
unabated while weeds shrivel and die. Bugs and diseases that have been the bane of farmers since time
immemorial leave the crops unscathed. Third, GM foods can be manipulated to withstand the rigors of longdistance shipping and to ripen more slowly. This allows food to reach more people in more locations, ensuring
variety in peoples diets.[GMOs] can provide farmers with disease-free planting materials and develop
crops that resist pests and disease, reducing use of chemicals that harm the environment and human
health. It can provide diagnostic tools and vaccines that help control devastating animal diseases. It can
improve the nutritional quality of staple foods such as rice and cassava and create new products for health and
industrial uses.genetic engineering the manipulation of an organisms genes. Same as gene splicing.
Genetic marker a known DNA sequence that can be copied and transferred to another organism in order
to replace a faulty DNA sequence.genetic modification intentional manipulation of an organisms
genesgenetics the science of heredity and variation in living organisms. genetic use restriction technology
(GURT) commonly called terminator technology, it refers to restricting genetically modified plants
(which are typically patented) by engineering the seeds second generation to be sterile. As of 2009, no such
seed is on the market anywhere in the world.genome the complete set of genes of a particular
species.genomics the study of an organisms entire genome, including DNA sequencing and gene mapping, as
opposed to the study of specific genes, which falls under the aegis of molecular biology.genotype the inherited
instructions contained in an organisms genetic code.germ-line genes genes that are included in the sex cells
and can be passed on to offspring.guanine one of the four bases in DNA and RNA. Guanine is always paired
with cytosine.HIV (Human Immunodeficiency Virus) a transmissible retrovirus that causes AIDS, acquired
immunodeficiency syndrome.horizontal gene transfer (HGT) the process of an organism incorporating
genetic information from an organism other than its parent. Significantly less common than vertical
gene transfer, in which genetic information is passed on through an organisms offspring, horizontal gene
transfer may be the mechanism by which GM seed contaminates nearby nonGM crops. Also called lateral
gene transfer.human genome the genome of Homo sapiens,which contains more than 3 billion DNA base pairs
and between 20,000 and 25,000 genes.Human Genome Project international research effort implemented by
the U.S. Department of Energy and the National Institutes of health in 1990 2 to identify the base pair
sequences and genes of the human genome. The project was completed in 2003 and has made its data
available to the public.hybrid the offspring resulting from the cross-breeding of plants or animals. Plant
hybrids are relatively common and have been cultivated for centuries. Animals hybrids between different
breeds of dogs, for example, are also common; but less common are hybrids between felines, such as the
liger, a cross between a lion and a tiger.induced pluripotent stem cell (iPS) a stem cell created from an adult
somatic cell (such as a skin cell) that is able to be transformed into any kind of cell (i.e., a pluripotent cell).
Unlike embryonic stem cell research, iPS research does not require the destruction of an embryo.infectious
disease a disease spread by pathogens of any kind, including bacteria, viruses, fungi, prions, and parasites,
that can be transmitted from one organism to another, as opposed to an inherited disease.inoculate
introducing a substance into the body for the purpose of boosting the bodys immune system to a certain
disease in vitro fertilization (IVF) a form of assisted reproductive technology in which a females egg and a
males sperm are extracted and fertilized outside the womb (in vitro). The resulting zygote is implanted in
the females uterus and the pregnancy continues normally.junk DNA the common term for the vast portions of
an organismsBiotechnology also has the potential to improve non-food uses of crops as sources of
industrial feedstocks or new materials such as biodegradable plastics. Plant-based materials can provide
both molecular building blocks and more complex molecules for the manufacturing, and energy and
pharmaceutical industries. Modifications under development include alterations to carbohydrates, oils, fats and
proteins, fibre and new polymer production. Under the appropriate economic and fiscal conditions,
biomass could contribute to alternative energy with both liquid and solid biofuels such as biodiesel and
bioethanol as well as processes such as bio-desulphurisation. Plant genomics also contributes to conventional
improvements through the use of marker-assisted breeding.

Genetic Modification of Plants

New traits introduced to crop
plants by genetic engineeringhave the potential to increase crop yields, improve agricultural practices, or add
nutritional quality to products. For example, transgenic crop plants capable of degrading weed killers allow
farmers to spray weeds without affecting yield. Use of herbicide-tolerant crops may also allow farmers to move
away from pre-emergent herbicides and reduce tillage, thereby decreasing soil erosion and water loss.
Transgenic plants that express insecticidal toxins resist attacks from insects. Crops engineered to resist insects
are an alternative to sprays, which may not reach all parts of the plant. They are also cost effective, reducing
the use of synthetic insecticides. Genetic engineering has also been used to increase the nutritional value of
food; "golden rice" is engineered to produce beta-carotene, for example. Edible vaccines, present in the plants
we eat, may be on the horizon.
The new traits expressed in such transgenic plants are derived from a variety
of other organisms. Scientists have given a gene from the bacterium
Salmonella to cultivars of soybeans, corn, canola, and cotton to degrade the
pesticide glyphosphate (Roundup TM). The gene for the insecticidal toxin in
transgenic cotton, potato, and corn plants comes from the bacterium Bacillus
thuringiensis (Bt). One of the genes allowing vitamin A production in golden
rice is derived from the bacterium Erwiniauredovora; others are from the
daffodil.

GENETIC ENGINEERING OF
PLANTS

Generating transgenic plants is key to introducing new crop traits and carrying out gene studies, both in discovery and applied
settings. The development of genetically modified plants often requires a complex design of DNA elements to achieve optimum
expression effects. We offer solutions for genome editing, cloning, DNA assembly, amplification and analysis, as well
as Agrobacterium tumefaciens LBA4404 for plant-cell transformation.

Plant Transformation
Agrobacterium-mediated transformation is the easiest, most simple, and widely used transformation technique in plant
biotechnology applications, and we offer A. tumefaciens LBA4404 cells and antibiotics to enable your research in this field.

Agrobacterium tumefaciens LBA4404 competent cells

Antibiotic to eliminate Agrobacterium after transformation: Carbenicillin

Reagents to select for transgenic plant cells after transformation:

o

Kanamycin

Basic information about plants

Plants can be self-fertilized.
Most plants can be classified into one of two classes,
which differ in many properties:
Monocotylenous(monocots)--seed have a single leaf (wheta,
rice, corn).
Dicotyledonous (dicots) - seed have two leaves.

Plant cells are surrounded by a cellulose cell wall

Whole plants can be regenerated from individual cultured
cells.

How to introduce foreign DNA into plants?

Bacterial infection (Agrobacterium tumefaciens)
Viral infections (tobacco mosaic virus)
Gunshot introduction

Agrobacterium infection causes crown gall

tumors in plants
Plants need to have a

wound
A piece of DNA
(calledT-DNA) from a
plasmid (the Ti
plasmid) in the
Agrobacterium gets
incorporated into theplant cell genome

Instead of the Ti plasmid, a synthetic binary

vector system can be used for gene transfer
Small plasmid containing
gene of interest flanked by
border regions.
Larger plasmid containing vir
genes.
Agrobacterium can be
transformed with both
plasmids.
Advantage; easy to generate
small plasmids with cloned

For monocots, agrobacterium infection not

effective--introduce genes using gene gun

GENETIC ENGINEERING OF BACTERIA

Genetically modified bacteria were the first organisms to be modified in the laboratory, due to their
simple genetics.[1]These organisms are now used for several purposes, and are particularly important in producing large
amounts of pure human proteins for use in medicine.[2]
The first example of this occurred in 1978 when Herbert Boyer, working at a University of California laboratory, took a
version of the human insulin gene and inserted into the bacterium Escherichia coli to produce synthetic "human" insulin.
Four years later, it was approved by the U.S. Food and Drug Administration.
The drug industry has made use of this discovery in its quest to produce medication for diabetes. Similar bacteria have
been used to produce clotting factors to treat haemophilia,[and human growth hormone to treat various forms ofdwarfism.
[

These recombinant proteins are safer than the products they replaced, since the older products were purified

from cadavers and could transmit diseases.Indeed, the human-derived proteins caused many cases
of AIDS and hepatitis C in haemophilliacs and Creutzfeldt-Jakob disease from human growth hormone.
For instance, the bacteria which cause tooth decay are called Streptococcus mutans. These bacteria consume leftover
sugars in the mouth, producing lactic acid that corrodes tooth enamel and ultimately causes cavities. Scientists have
recently modified S. mutans to produce no lactic acid.These transgenic bacteria, if properly colonized in a person's
mouth, could reduce the formation of cavities. Transgenic microbes have also been used in recent research to kill or
hinder tumors, and to fight Crohn's disease.Genetically modified bacteria are also used in some soils to facilitate crop
growth, and can also produce chemicals toxic to crop pests.
GM bacteria have also been developed to leach copper from ore,clean up mercury pollution [13] and detect arsenic in
drinking water.