VIROLOGY

UNIT 6. Use of bacteriophages in gene cloning

Virology Dr.Mebrate Dufera

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 Bacteriophage
 A bacteriophage is a type of virus that destroys bacteria.
 bacteriophages, which use bacteria as their hosts.
 It consists of
 a head, containing the genetic material,
 Neck and
 a tail, which attaches to the exterior of a bacterium.
 The genetic material of the bacteriophage passes from its head
through its tail into the bacterium.
 The genetic material then directs the bacterium to create new
bacteriophages,
 which eventually burst from their host and, in the process,
destroy the bacterium.
 The released bacteriophages
Virology
attack nearby bacteria, and the
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infection process continues. 2018
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 Gene cloning
 Gene cloning is a method for making many identical copies of
a gene by inserting the gene into a living host cell.
 Under appropriate conditions, the host cell will replicate the
“foreign” DNA along with its own DNA whenever it divides.
 With each cell division, the number of copies of the “foreign”
gene doubles.
 This technique permits biologists to produce large quantities
of a single gene so that they can study the gene in detail.
 For example, scientists can produce proteins that are used as
enzymes in commercial food processing techniques or
proteins that can be used to treat disease.

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 Vectors
 Vectors are tools used by molecular biologists to insert genes or
pieces of foreign DNA into host cells.
 Two naturally occurring vectors that are used to insert foreign
DNA into bacterial cells are bacteriophages and plasmids.
 Bacteriophages (or phages, for short) are viruses that infect
bacterial cells by injecting their genetic material into the
bacterial cell.
 Plasmids are small circles of DNA that are sometimes present in
bacteria in addition to the larger circle of DNA that constitutes
the main bacterial genome.
 When DNA from 2 different sources is combined, the resulting
DNA is called recombinant DNA. If recombinant DNA gets inside
a host cell, it can replicate along with the DNA of the host cell.

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 Non-replicating viruses as vectors in gene therapy and for
development of new types of vaccine
Use of viruses to produce recombinant proteins in animals and Plants

 The use of plants for vaccine production is emerging as an attractive

alternative to traditional production systems.
 Plant expression systems have several advantages such as
 the absence of risk of contamination with animal pathogens,
 potentially low production costs, and
 the possibility of very large-scale production.
 They also offer the option of producing edible vaccines.
 These recombinant proteins have a wide range of important applications,
including
 vaccines and therapeutics for human and animal health,
 industrial enzymes,
 new materials and components of novel nano-particles for various
applications.
 The majority of recombinant proteins are produced by traditional biological
"factories," that is, predominantly mammalian and microbial cell cultures
along with yeast and insect cells.
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 UNIT 7. Emerging viruses & future trends HIV Bird or
Avian virus, Ebola virus
 In humans, such infections will more often be zoonotic
 Zoonotic: the transmission of a virus from wild or domesticated animals to
humans with attendant disease.
 As we discover new viruses and relate new and well-known viruses to
specific diseases, we can anticipate public pressure to rapidly develop
antivirals and vaccines.
 For example, at the start of the 21st century, we witnessed several
examples of emerging infections followed by exaggerated (but not
necessarily unwarranted) public reactions.
 The global concern and almost immediate response of scientists and
health officials to the SARS and West Nile virus epidemics are cases in
point.
SARS ? Severe acute respiratory syndrome (SARS) is a viral
respiratory disease of zoonotic origin caused by the SARS
coronavirus (SARS-CoV)
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 What will tomorrow bring?
 How will we deal with these new infections?
• Improved surveillance,
• more-rapid re- agent sharing and information transfer,
• more-effective quarantine procedures, and
• various public health measures will undoubtedly contribute to
controlling emerging diseases,
• but increasing attention and resources are likely to be
devoted to maintaining,
• as well as expanding, the roster of antivirals and vaccines.

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 Co-infection of the same cell with different related viruses
allows for genetic reassortment (exchange of genome
segments) and intramolecular recombination.
 Genetic alterations can alter virulence or allow viruses to gain
access to new cell types or new animal hosts.
 Many scientists believe that HIV is derived from a closely
related monkey virus, SIV (simian immunodeficiency virus),
that acquired the ability to infect humans.
 Many of today’s emerging viruses may have similar histories.

Virulence? virulence refers to the degree of

damage caused by a microbe to its host.

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 MEDICALLY VERY IMPORTANT VIRUSES
A. AIDS Virus  HIV can be transmitted:
• The human sexually, through contact
immunodeficiency virus with contaminated blood,
(HIV), which causes tissue, or needles, and from
acquired immunodeficiency mother to child during birth
syndrome (AIDS), or breast-feeding.
• principally attacks T-4
lymphocytes CD4), a vital
part of the human immune
system.
• As a result, the body’s
ability to resist
opportunistic viral,
bacterial, fungal, protozoal,
and other infection is
greatly weakened.

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 How the HIV Infection Cycle Works
What factors influence the time period from initial HIV
infection and the development of symptoms?
 HIV is an unusual viral disease in that individuals can carry the virus
for very extended periods of time without becoming ill, or
exhibiting any symptoms at all.
 However, at any time, the symptoms of AIDS can manifest
themselves.
 The variation in time until onset of the disease is associated with
several factors. The most important is
 having a well-functioning immune system.
 Individuals with weak immune systems, such as older people or
people with certain types of pre-existing infections develop AIDS
more rapidly, on average, than other people do.
 Other factors that influence the onset of disease are treatment with
various medications, the strain of HIV involved, and the genotype of
the infected person.

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 The first contact between a virus particle and its host cell
occurs:
 when an outer viral structure docks with a specific molecule
on the cell surface.
 For example, a glycoprotein called gp120 on the surface of
the human immunodeficiency virus specifically binds to the
CD4 molecule T lymphocytes.

 Most cells that do not have surface CD4 molecules generally

cannot be infected by HIV.
Eg. Wife +ve ; Husband –ve or
Husband –ve; Wife +ve?????
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 Life cycle of Human Immunodeficiency Virus (HIV)

 Pass through the following 8 steps:

1. Free-floating HIV
2. HIV Binds with Cell-----Adsorption
3. HIV Infects Cell-----------Entry
4. Viral RNA Converted to DNA-Reverse transcription
5. Viral DNA Spliced into Cellular DNA
6. Transcriptions----mRNA-----DNA
7. Assembly
8. Budding
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 1. Free-floating HIV
 Like all viruses, human
immunodeficiency virus
(HIV) is comprised of only
genetic material, a few
proteins, and a protective
envelope.
 Its genetic material,
carried by single-
stranded RNA molecules,
contains all the
information necessary to
make more viruses.
 HIV can not reproduce
itself outside of a cell, but
when HIV invades a living
cell, it turns the cell into
a factory for making
more HIV.

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 2.HIV binds with cell
• HIV binds to
specific receptors
on the cell surface.
• These receptors
are found on only
a few types of
cells, including
certain white
blood cells,
intestinal cells, and
brain cells.

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 3.HIV Infects cell

• HIV penetrates
the cell
membrane and
releases its
contents into
the host cell.

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 4.Viral RNA converted to DNA

• A viral protein,
the enzyme
reverse
transcriptase,
converts single
strands of RNA
into double-
stranded DNA.

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 5.Viral DNA Spliced into Cellular DNA

• Another viral
protein, the
enzyme integrase,
splices the viral
DNA into the DNA
in the host cell's
nucleus.
• The cell becomes
unable to
distinguish viral
DNA from its own
genetic material
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 6.Transcription

• The host cell

carries out the
instructions
encoded in
the viral DNA,
making copies
of the viral
RNA and viral
proteins.

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 7.Assembly
• A third viral
protein, the
enyzme protease,
cleaves the viral
proteins into
functional
enzymes.
• The viral proteins
and RNA come
together in the cell
to facilitate the
production of a
new virus.
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 8.Budding
• The viral
proteins and
RNA are
packaged into a
newly formed
HIV, which
buds from the
host cell and is
soon ready to
infect other
cells.
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 B. Rabies Virus
• The rabies virus is usually transmitted to
humans by a bite from an infected dog, but
the bite of any animal (wild or domestic) is
suspect in an area where rabies is present.
• Symptoms of the disease appear after an
incubation period of ten days to one year and
include fever, breathing difficulties, and
muscle spasms in the throat that make
drinking painful.
• Death almost invariably occurs within three
days to three weeks of the onset of
symptoms.
• For this reason, the emphasis of treatment is
on prevention.
Notice:
Incubation period: the period between infection
and Symptom.
Prepatent period: the period between infection
and Diagnosis (parasite detection)

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 C. Hepatitis B Virus
• The hepatitis B virus (HBV)
causes inflammation of the
liver.
• The virus is recognizable
under magnification by the
round, infectious “Dane
particles” accompanied by
tube-shaped, empty viral
envelopes.
• Symptoms of hepatitis B
infection include jaundice
and a flulike illness, while
chronic infection can lead to
serious problems such as
cirrhosis and cancer of the
liver.
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 D. Smallpox Viruses
(Eradicated)
• A colored-transmission
electron micrograph
shows a group of
Orthopoxvirus variola,
the virus that causes
smallpox.
• Once greatly feared for
its ability to kill or disable
its victims, smallpox was
eradicated by 1979
through a worldwide
vaccination campaign.
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 E. Ebola Virus Infection
• The Ebola virus is highly
infectious and can spread
through the use of unsterilized
needles or through contact with
an infected individual or the
corpse of someone who has
died from the disease.
• About one week after infection,
the virus begins attacking blood
and liver cells (1).
• As the disease swiftly
progresses, the virus may
destroy vital organs such as the
liver and kidneys (2),
• leading to massive internal
bleeding (3).
• Shock and respiratory arrest
soon follow, then death.
In: Congo- Vaccination started !!

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