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Benefits of

Gene
Technology
Angga Sendiputra
Christian Chandra

The Role of Recombinant DNA


Technology in Biotechnology

Recombinant DNA Technology


Intentional modification of organisms
genomes for practical purposes
Three goals
Eliminate undesirable phenotypic traits
Combine beneficial traits of two or more
organisms
Create organisms that synthesize
products humans need

The Tools of Recombinant DNA


Technology

Mutagens
Physical and chemical agents that
produce mutations
Scientists utilize mutagens to
Create changes in microbes genomes to
change phenotypes
Select for and culture cells with beneficial
characteristics

The Tools of Recombinant DNA


Technology

The Use of Reverse Transcriptase to Synthesize


cDNA
Isolated from retroviruses
Uses RNA template to transcribe molecule
of cDNA
Easier to isolate mRNA molecule for desired
protein first
mRNA of eukaryotes has introns removed
Allows cloning in prokaryotic cells

The Tools of Recombinant DNA


Technology

Synthetic Nucleic Acids


Molecules of DNA and RNA produced in
cell-free solutions
Uses of synthetic nucleic acids
Elucidating the genetic code
Creating genes for specific proteins
Synthesizing DNA and RNA probes to
locate specific sequences of nucleotides
Synthesizing antisense nucleic acid
molecules

The Tools of Recombinant DNA


Technology
Restriction

Enzymes
Bacterial enzymes that cut DNA
molecules only at restriction sites
Categorized into two groups based
on type of cut
Cuts with sticky ends
Cuts with blunt ends

The Tools of Recombinant DNA


Technology

Vectors
Nucleic acid molecules that deliver a gene
into a cell
Useful properties
Small enough to manipulate in a lab
Survive inside cells
Contain recognizable genetic marker
Ensure genetic expression of gene
Include viral genomes and plasmids

The Tools of Recombinant DNA


Technology
Gene

Libraries
A collection of bacterial or phage
clones
Each clone in library often contains
one gene of an organisms genome
Library may contain all genes of a single
chromosome
Library may contain set of cDNA
complementary to mRNA

Pharmaceutical and
Therapeutic Applications
Protein

Synthesis
Vaccines
Genetic Screening
DNA Fingerprinting
Gene Therapy
Medical Diagnosis
Xenotransplants

Pharmaceutical and
Therapeutic Applications

Protein Synthesis

Scientists have inserted


synthetic genes for insulin,
interferon, and other proteins to
bacteria and yeast cells so that
the microbes synthesize these
proteins in vast quantities.

Pharmaceutical and
Therapeutic Applications
Vaccines

Scientists now use recombinant DNA


technology to produce safer vaccines.
After they insert the genes that codes for
a phatogens antigens to a vector, they
inject recombinant vector or the proteins
it produces into a patient. Thus the
patients immune is exposed to a subunit
of pathogen, not the pathogen itself.

Pharmaceutical and
Therapeutic Applications
Genetic

Screening
Genetic screening, also known as
DNA testing, allows the genetic diagnosis
of vulnerabilities to inherited diseases,
and can also be used to determine a
child's parentage (genetic mother and
father) or in general a person's ancestry.
Genetic testing identifies changes in
chromosomes, genes, or proteins

Pharmaceutical and
Therapeutic Applications
DNA

Fingerprinting
DNA profiling (also called DNA testing,
DNA typing, or genetic fingerprinting) is a
technique employed by forensic
scientists to assist in the identification of
individuals by their respective DNA
profiles. DNA profiles are encrypted sets
of letters that reflect a person's DNA
makeup, which can also be used as the
person's identifier.

Pharmaceutical and
Therapeutic Applications
Gene

Therapy
An existing use of recombinant DNA
technology is gene therapy, in which
missing or defective genes are replaced
with normal copies. A researcher could
remove a few defective genetically
defective cells from a patient, insert
normal genes, and replace the cells into
patient.

Pharmaceutical and
Therapeutic Applications
Medical

Diagnosis
Clinical microbiologists use PCR,
fluorescent genetic probes, and DNA
microarrays in diagnostic applications.
They examine specimens from patients
for the presence of viruses or bacterial
phatogens.

Pharmaceutical and
Therapeutic Applications
Xenotransplants

Xenotransplants are animal cells,


tissues, or organs introduced into the
human body. Because of the existence
of recombinant DNA technology, it is
theoretically feasible to insert functional
human genes into animals to direct them
to produce organs and tissues for
transplantation into human.

Agricultural Applications
Herbicide

Resistance
Salt Tolerance
Freeze Resistance
Pest Resistance
Improvements in Nutritional Value and
Yield

Agricultural Applications
Herbicide

Resistance
One of the most famous kinds of GM
crops is glyphosate-resistant trait.
Glyphosate, kills plants by interfering with
the shikimate pathway in plants, which is
essential for the synthesis of the aromatic
amino acids. Glyphosate inhibits the
enzyme 5-enolpyruvylshikimate-3phosphate synthase (EPSPS).

Agricultural Applications
Herbicide

Resistance (cont.)
Some micro-organisms have a version
of EPSPS that is resistant to glyphosate
inhibition. One of these was isolated from
an Agrobacterium strain CP4 (CP4 EPSPS)
that was resistant to glyphosate. This CP4
EPSPS gene was cloned and transfected
into soybeans.

Agricultural Applications
Salt

Tolerance
Scientists have successfully removed
the gene for salt tolerance from the
inedible salt-tolerant plants and inserted
it to tomato and canola plants to create
food crops that can grow in salty soil. The
transgenic plant can also remove salt
from soil and make it suitable to
unmodified crops as well.

Agricultural Applications

Freeze resistance
Abiotic stress like frost is limiting factor to
the growth of tomatoes. While no genetically
modified stress tolerant plants are currently
commercialised, transgenic approaches
have been researched. An early tomato was
developed that contained an antifreeze
gene (afa3) from the winter flounder with the
aim of increasing the tomato's tolerance to
frost.

Agricultural Applications

Pest Resistance

Tomatoes resistant to a root knot


nematode have been created by inserting a
cysteine proteinase inhibitor gene from taro.
A chemically synthesised ceropin B gene,
usually found in the giant silk moth
(Hyalophora cecropia), has been introduced
into tomato plants and in vivo studies show
significant resistance to bacterial wilt and
bacterial spot.

Agricultural Applications
Improvements

in Nutritional Value and

Yield
Recently scientists have increased the
production of anthocyanin, an
antioxidant in tomatoes in several ways.
One group added a transcription factor
for the production of anthocyanin from
Arabidopsis thaliana, whereas another
used transcription factors from
snapdragon (Antirrhinum)

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