CHAPTER 4: Lesson 15: Introduction to Genetic Engineering and Molecular Methods Flashcards
is the deliberate modification of an organism’s genetic
information by directly changing its nucleic acid genome and is achieved by a group
of methods known as recombinant DNA technology.
Genetic engineering
a group of methods known as
recombinant DNA technology
is of value on basic research on gene structure and
function, production of useful proteins by novel methods, generation of transgenic
plants and animals, medical diagnosis and treatment or genome analysis by DNA
sequencing.
Genetic manipulation
The term has been coined to describe the
ethical problems that exist in modern genetics
genethics
are likely to increase in both
number and complexity as genetic engineering technology becomes more
sophisticated
genethics
example of advanced genetic engineering technology
development of Genetically Modified Organisms (GMOs).
The process of artificially
introducing foreign genes into organisms is termed
transfection
The process of artificially
introducing foreign genes into organisms is termed transfection, and the recombinant
organisms produced in this way are called
transgenic or Genetically Modified
organisms (GMOs)
Foreign genes have been inserted into a variety of microbes, plants, and
animals through
recombinant DNA techniques
One useful property of DNA
it readily
anneals (changes its binding properties in response to heating and cooling)
Exposure to temperatures _____________ causes DNA to become
temporarily denatured
just below boiling (90–95°C)
Exposure to temperatures just below boiling (90–95°C) causes DNA to become
_______.
temporarily denatured
When heating is followed by gradual cooling, two single DNA strands _________ by hydrogen bonds at complementary sites
rejoin (renature)
When heating is followed by gradual cooling, two single DNA strands rejoin (renature) by _________ at complementary sites
hydrogen bonds
is a necessary feature of the polymerase chain reaction (PCR) and nucleic acid probes
Annealing
Annealing is a necessary feature of the __________.
Polymerase Chain Reaction (PCR)
&
Nucleic Acid Probes
possessed by bacteria which allows them to recognize
specific regions of DNA for cleavage in producing DNA fragments to be used in
genetic engineering
Restriction enzymes
DNA sequences recognized by restriction enzymes are
predominantly __________.
palindromes
Inverted sequence repetitions
palindromes
Methods used to Size, Synthesize, and Sequence DNA
Hybridization
DNA sequencing
Nucleic acid probes:
It is the process of combining two complementary single-stranded
DNA or RNA molecules and allowing them to form a single double-stranded
molecule through base pairing.
Hybridization
Types of hybridization
- Northern blot
- Southern blot
- Western blot
Hybridization of DNA to RNA, which
provides quantitative information about RNA synthesis.
Northern blot
Hybridization of DNA to DNA.
Southern blot
This method is useful to detect specific DNA sequences in restriction fragments
separated on gels. These blots can be used to detect overlapping restriction
fragments.
Southern blot
Southern blot is used to….
detect overlapping restriction
fragments
It is a technique used for detection of genes, in which antibodies
are used to detect cloned genes by binding to their protein products.
Western blot
are used to detect cloned genes by binding to their protein products
antibodies
It shows gene structure that helps research
workers to find out the structure of gene products.
DNA sequencing
DNA Sequencing Applications
■ Information obtained by DNA sequencing makes it possible to understand or
alter the function of genes.
■ DNA sequence analysis demonstrates regulatory regions that control gene
expression and genetic “hot spots” particularly susceptible to mutation.
■ Comparison of DNA sequences shows evolutionary relationships that provide
a framework for definite classification of microorganisms including viruses.
■ Comparison of DNA sequences facilitates identification of conserved regions,
which are useful for development of specific hybridization probes to detect
microorganisms including viruses in clinical samples
are segments of DNA and RNA labeled with
radioisotopes or enzymes that can hybridize to complementary nucleic acids with
high degree of specificity.
Nucleic acid probes
The basis of specially formulated oligonucleotide tracers called
gene probes
A molecule labeled with a radioactive isotope, dye, or enzyme that is used to locate particular sequence or gene on a DNA molecule
DNA Probe
DNA probe is also known as
hybridization probe
hybridization probe is also known as
DNA Probe
A sequence that is used to locate a specific DNA/RNA fragment to take to PCR to be replicated.
DNA Probe
a technique that syntheysize large quantities of a DNA fragment without cloning it.
Polymerase Chain Reaction (PCR)
By this technique, large quantities of a particular DNA sequence can be prepared
Polymerase Chain Reaction (PCR)
This method can generate tens of billions of copies of a particular
DNA fragment (the sequence of interest, DNA of interest, or target DNA) from a DNA
extract (DNA template)
Polymerase Chain Reaction (PCR)
Three Major Steps Involved in the PCR Technique:
1) denaturation
2) annealing
3) extension
Steps Involved in the PCR Technique
STEP 1; the DNA is denatured at high
temperatures(from 90 - 97 degrees Celsius).
STEP 2: primers anneal to the DNA template strands to prime extension.
STEP 3: extension occurs at the end of the
annealed primers to create a complementary copy strand of DNA.
DNA is denatured in what temperatures?
90 - 97°C
This step in the PCR cycle effectively
doubles the DNA quantity
STEP 3: Extension
Denaturation temperature
94°C
Annealing temperature
54°C
Extension temperature
72°C
A PCR can either be what?
Qualitative or Quantitative
is also referred to real time PCR
Quantitative PCR techniques
It gives an idea about how much DNA amount
present in the sample
Quantitative PCR techniques
Is used for detecting a
specific DNA segment
Qualitative PCR techniques
is another modification of PCR in which two or more target sequences can be demonstrated simultaneously in a single specimen at the same time
Multiplex PCR
is designed to collect data as the reaction is proceeding,
which is more accurate for DNA and RNA quantitation and does not require
laborious post PCR methods.
Real-time PCR
A technique mainly used to change the
phenotype of an organism (host) when a genetically altered vector is introduced and integrated into the genome of the organism
Recombinant DNA Technology
Basically, this process introduce
a foreign piece of DNA structure into the genome of the host. This particular
gene that is introduced is referred to as the recombinant gene.
Recombinant DNA Technology
This particular
gene that is introduced in Recombinant DNA Technology is referred to as the
recombinant gene
Biological Products of Recombinant DNA Technology
Genetically Modified Organisms (GMOs)
Gene Therapy
The process of artificially introducing foreign genes into organisms is termed
transfection
are available for a variety of biotechnological applications. Because they are unique life forms that would never have otherwise
occurred, they can be patented.
Transgenic “designer” organisms
is a technique for replacing a faulty gene with a normal one in
individuals with fatal or extremely debilitating genetic diseases.
Gene therapy
The inherent benefit
of this therapy is to permanently cure the physiological dysfunction by repairing the
genetic defect
Gene therapy
