CHAPTER 27 BIOTECHNOLOGY - Garland Science

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Microbiology: A Clinical Approach
[9780815365143] © Garland
Science
ISBN: 978
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6514
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Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
CHAPTER 27
BIOTECHNOLOGY
© David Parker / Science Photo Library
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Microbiology: A Clinical Approach,
by Tony
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© Garland Science
Microbiology: A Clinical Approach
© Garland Science
WHY IS THIS IMPORTANT?

Biotechnology has made routine what used to
be fantastic.

Advances using biotechnology already impact
the health care community.

Health care professionals should have a basic
understanding of this field.
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Microbiology: A Clinical Approach,
by Tony
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© Garland Science
Microbiology: A Clinical Approach
© Garland Science
OVERVIEW
Microbiology: A Clinical Approach
[9780815365143] © Garland
Science
ISBN: 978
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Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
OVERVIEW

Biotechnology is ubiquitous.

Affects every aspect from food to reproduction to
politics

Used in everything from manufacturing to
bioweapons
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Microbiology: A Clinical Approach,
by Tony
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© Garland Science
Microbiology: A Clinical Approach
© Garland Science
OVERVIEW

Development of the microscope in the 1700s
changed science.

Scientists could see cells and identify some of their
functions.

Disease mechanisms could be studied at the cellular
level.

300 years later we have:

Electron microscopy

Genetic engineering

Molecular biology
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Microbiology: A Clinical Approach,
by Tony
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© Garland Science
Microbiology: A Clinical Approach
© Garland Science
BIOTECHNOLOGY DEFINED

The field devoted to applying techniques of
biochemistry, cell biology, biophysics, and
molecular biology to addressing issues related
to human beings and the environment in which
they live.
Microbiology: A Clinical Approach
[9780815365143] © Garland
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ISBN: 978
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Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
BIOTECHNOLOGY DEFINED

Despite the diversity of cell types in nature most
cells have the same basic properties.

Provide opportunities for external manipulation

DNA and RNA are the universal genetic
materials.

All living things use DNA and RNA in some form.

Information can be shared or exchanged by different
organisms.

Can create specific proteins in a myriad of different
organisms

This is the heart of biotechnology.
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Microbiology: A Clinical Approach,
by Tony
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© Garland Science
Microbiology: A Clinical Approach
© Garland Science
BIOTECHNOLOGY INDUSTRY

The industry has grown explosively over the past 20
years.

Annual revenues are well over $40 billion.

About 1500 companies in the US alone employing over 188,000
people.

More than 200 diseases currently targeted for treatment
with drugs produced using biotechnology.

Alzheimer’s disease

Cancer

Multiple sclerosis

Diabetes

Arthritis
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Microbiology: A Clinical Approach,
by Tony
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© Garland Science
Microbiology: A Clinical Approach
© Garland Science
BIOTECHNOLOGY INDUSTRY

More than 190 new medicines already
developed.

Approximately 30 more new medicines are
produced each year

Process of discovery of new drugs very
expensive and time consuming.

Takes up to 16 years
Microbiology: A Clinical Approach
[9780815365143] © Garland
Science
ISBN: 978
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6514
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Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
BIOTECHNOLOGY INDUSTRY
ISBN: 978
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6514
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Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
BIOTECHNOLOGY INDUSTRY

In the US biotechnology is regulated by the FDA,
USDA, and EPA.

Biotechnology products are already in use around
the world.

Genetically engineered food

Hundreds of medical diagnostic tests and treatments

Environmentally sound hazardous waste disposal

More efficient ways to use water and energy
ISBN: 978
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6514
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Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
HISTORY OF BIOTECHNOLOGY

Actually used as early as 2000
-
4000BC

Humans learned to use plants and plant by
-
products
medicinally and in cooking

1590s

Lens maker Janssen invents the compound
microscope.

1663

Hooke observes cell structure in plants and
animal tissue.

1675

van Leeuwenhoek observes individual
bacteria.

Late 1700s

Jenner uses cowpox virus to vaccinate
against smallpox.

Mid 1800s

explosion in studies on microorganisms
and molecules in cells.
Microbiology: A Clinical Approach
[9780815365143] © Garland
Science
ISBN: 978
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Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
HISTORY OF BIOTECHNOLOGY

1870
-
1890

Koch developed his postulates.

1928

Fleming discovers penicillin.

1946

recombination of viruses.

1953

Watson and Crick discover the structure of
DNA.

1966

human genetic code is cracked.

1970s

recombinant DNA experimentation

Gene targeting

Gene splicing
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Microbiology: A Clinical Approach,
by Tony
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© Garland Science
Microbiology: A Clinical Approach
© Garland Science
HISTORY OF BIOTECHNOLOGY

1980s

human genes placed into bacteria.

Human insulin first success

1980s

DNA fingerprinting is widely accepted
legally.

1980s

EPA approves first transgenic crop (tobacco).

1980s

first recombinant vaccine approved (for
Hepatitis B).

1980s

beginning of use for bioremediation.

First used on oil spills.
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Microbiology: A Clinical Approach,
by Tony
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© Garland Science
Microbiology: A Clinical Approach
© Garland Science
HISTORY OF BIOTECHNOLOGY

1990s

beginning of the human genome project.

1990s

first transgenic cow created.

First sheep cloned from an adult sheep cell.

More animals are cloned.

1990s

human embryonic stem cell lines are
established.

2002

human genome project is completed and
published.

2004

cloning of embryonic stem cells.
Microbiology: A Clinical Approach
[9780815365143] © Garland
Science
ISBN: 978
-
0
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8153
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6514
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Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
TOOLS

Tools most often used in biotechnology:

Cell culture

Genetic engineering

Monoclonal antibodies

Recombinant DNA
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Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
TOOLS
ISBN: 978
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Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
CELL CULTURE

Growing cells outside the organism is a routine
and necessary technique.

Enzymes are used to isolate cells from tissues.

Cells are cultured in growth media.

Cell lines are used to harvest useful products
made by particular cells.

Some cell types are easy to grow.

Fibroblasts and plant cells
Microbiology: A Clinical Approach
[9780815365143] © Garland
Science
ISBN: 978
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0
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6514
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Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
CELL CULTURE

Some cell types are more difficult to grow.

Some cell types cannot be grown in culture.

Plant cell lines produce flavors for food,
emulsifiers, and thickening agents.

Insect cell lines used to identify and improve
biocontrol agents for insect pests.
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Microbiology: A Clinical Approach,
by Tony
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© Garland Science
Microbiology: A Clinical Approach
© Garland Science
MAMMALIAN CELL CULTURES

Mammalian cell cultures are widely used in
biology.

Growth and/or identification of bacterial and viral
organisms

Studying the cell cycle

Fertility issues

Diagnosing disease

Provides a renewable source of DNA

Understanding the effects of drugs
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Microbiology: A Clinical Approach,
by Tony
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© Garland Science
Microbiology: A Clinical Approach
© Garland Science
VIRAL CELL CULTURES

Viruses can be grown in different types of cell
line.

Primary

derived directly from a tissue sample

Continuous

transformed cells that grow
continuously in culture

Diploid

developed from human embryos

Can only survive about 100 generations (doublings)

Used to grow viruses requiring a human host
Microbiology: A Clinical Approach
[9780815365143] © Garland
Science
ISBN: 978
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8153
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6514
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3
Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
STEM CELLS

Stem cells are unspecialized.

Can divide many times and remain unspecialized

Stem cells can be influenced to change into specific
cells

differentiation.

Through the use of chemical signals and
microenvironmental
factors

Ability to differentiate into variable types of cells is
referred to as plasticity

These cells are useful therapeutically or in research
trials.

Used to grow certain viruses
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Microbiology: A Clinical Approach,
by Tony
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© Garland Science
Microbiology: A Clinical Approach
© Garland Science
STEM CELLS
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Microbiology: A Clinical Approach,
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© Garland Science
Microbiology: A Clinical Approach
© Garland Science
STEM CELLS:
Adult

Populations of undifferentiated cells

Reside in various tissues e.g. brain, heart, bone
marrow, liver, and skin

Might be useful for patients requiring new tissue
or organs

Using patient’s own cells = no rejection

Some adult stem cells capable of
pluripotency

Able to differentiate into tissues other than that from
which they came
Microbiology: A Clinical Approach
[9780815365143] © Garland
Science
ISBN: 978
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6514
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Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
STEM CELLS:
Embryonic

Derived from fertilized eggs
in vitro.

Completely
pluripotent

Can become any cell type

Ideal for cell based therapy

Easily grown in the lab

Maintain undifferentiated state through numerous
generations

Can be differentiated into specific cell types
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Microbiology: A Clinical Approach,
by Tony
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© Garland Science
Microbiology: A Clinical Approach
© Garland Science
STEM CELLS:
Embryonic
ISBN: 978
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Microbiology: A Clinical Approach,
by Tony
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© Garland Science
Microbiology: A Clinical Approach
© Garland Science
STEM CELLS:
Embryonic
Microbiology: A Clinical Approach
[9780815365143] © Garland
Science
ISBN: 978
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6514
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Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
GENETIC ENGINEERING

Several distinct tools involved:

Monoclonal antibodies

Recombinant DNA technology
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Microbiology: A Clinical Approach,
by Tony
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© Garland Science
Microbiology: A Clinical Approach
© Garland Science
MONOCLONAL ANTIBODIES

Kohler and Milstein developed a technique for
constructing monoclonal antibodies.

Used immune cells hybridized with transformed
cells

Produced clone cells

Clone cells produced identical antibodies

monoclonal antibodies
ISBN: 978
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8153
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6514
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3
Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
MONOCLONAL ANTIBODIES
Microbiology: A Clinical Approach
[9780815365143] © Garland
Science
ISBN: 978
-
0
-
8153
-
6514
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3
Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
MONOCLONAL ANTIBODIES
ISBN: 978
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8153
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6514
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3
Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
MONOCLONAL ANTIBODIES

Monoclonal antibodies used for:

Diagnostic tests

Detecting harmful microorganisms in food

Locating environmental pollutants

Targeted delivery of chemotherapeutic drugs to
cancer cells

Inhibiting certain types of immune reactions to
protect transplants
ISBN: 978
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6514
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Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
MONOCLONAL ANTIBODIES

Humane concerns about using mice to produce
monoclonal antibodies:

Hollow fiber systems for tissue cultures

Genes for antibody can be cloned and put into high
production eukaryotic cells
Microbiology: A Clinical Approach
[9780815365143] © Garland
Science
ISBN: 978
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8153
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6514
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3
Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
RECOMBINANT DNA TECHNOLOGY

DNA molecule can be cut by enzymes and
recombined with other DNA molecules.

Foundation of recombinant technology.

Two frequently used techniques for replicating
DNA.

Cloning using a vector

Polymerase chain reaction
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Microbiology: A Clinical Approach,
by Tony
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© Garland Science
Microbiology: A Clinical Approach
© Garland Science
RECOMBINANT DNA
TECHNOLOGY

Vector method:

Gene is inserted into a vector, usually a plasmid,
using restriction enzymes.

Plasmid is transferred into another cell, usually a
bacterium.

Plasmid and inserted DNA replicates.

Vector method is restricted by the size of the
insert.
ISBN: 978
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6514
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Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
RECOMBINANT DNA
TECHNOLOGY
Microbiology: A Clinical Approach
[9780815365143] © Garland
Science
ISBN: 978
-
0
-
8153
-
6514
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3
Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
RECOMBINANT DNA
TECHNOLOGY
ISBN: 978
-
0
-
8153
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6514
-
3
Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
RECOMBINANT DNA
TECHNOLOGY

Polymerase chain reaction:

Amplifies the number of copies of the same gene

DNA strands are separated and used as templates
to make additional copies.

Copies can be inserted into other DNA molecules
using the vector method.
ISBN: 978
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6514
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Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
RECOMBINANT DNA
TECHNOLOGY

Recombinant DNA technology has many
applications.

Mass production of products like insulin to treat
diseases

Improved control of viral diseases

Treatment of genetic diseases
Microbiology: A Clinical Approach
[9780815365143] © Garland
Science
ISBN: 978
-
0
-
8153
-
6514
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3
Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
RECOMBINANT DNA
TECHNOLOGY
ISBN: 978
-
0
-
8153
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6514
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3
Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
RECOMBINANT DNA
TECHNOLOGY

Recombinant DNA technology has many
applications:

Improvement of food’s nutritional value

Decrease in environmental pollutants

Enhancement of agricultural biocontrol agents
ISBN: 978
-
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8153
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6514
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3
Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
RECOMBINANT DNA
TECHNOLOGY
Microbiology: A Clinical Approach
[9780815365143] © Garland
Science
ISBN: 978
-
0
-
8153
-
6514
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3
Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
RECOMBINANT DNA
TECHNOLOGY
© Sinclair Stammers / Science Photo Library
ISBN: 978
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6514
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Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
PROTEOMICS, GENOMICS, AND
BIOINFORMATICS

Genomics

determining all the genetic material
in a cell

Proteomics

determining all the proteins
expressed in a cell and their relationship to host
and other proteins

Genomics and proteomics generate large amounts
of information.

Information has to be analyzed by computer

bioinformatics.

Protein Data Bank (PDB), now called the Molecular
Modeling Database (MMDB)
ISBN: 978
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Microbiology: A Clinical Approach,
by Tony
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© Garland Science
Microbiology: A Clinical Approach
© Garland Science
MICROARRAY ANALYSIS

Technique used to study genes in DNA samples.

Small samples

Samples with large number of unknown genes

Comparing healthy and diseased tissue samples

Allows for cataloguing and analyzing hundreds of
thousands of genes at once.

Improves ability to predict which proteins are
produced by a given nucleic acid sequence.

Yields information on which sequences are
nonsense and can be discarded.
Microbiology: A Clinical Approach
[9780815365143] © Garland
Science
ISBN: 978
-
0
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8153
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6514
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3
Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
FORENSIC SCIENCE

DNA technology has become a fundamental
part of forensic science.

DNA fingerprinting routinely used for:

Paternity testing

Criminal prosecution
ISBN: 978
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Microbiology: A Clinical Approach,
by Tony
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© Garland Science
Microbiology: A Clinical Approach
© Garland Science
FORENSIC SCIENCE
© David Parker / Science Photo Library.
ISBN: 978
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6514
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3
Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
INDUSTRY AND APPLICATIONS

Industry is using products coming from
biotechnology.

In health care:

Early detection of diseases

Diagnostics and therapeutics
Microbiology: A Clinical Approach
[9780815365143] © Garland
Science
ISBN: 978
-
0
-
8153
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6514
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3
Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
INDUSTRY AND APPLICATIONS

Industry is using products coming from
biotechnology.

In pharmaceuticals:

New enzymes to streamline production processes
ISBN: 978
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0
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8153
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6514
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3
Microbiology: A Clinical Approach,
by Tony
Srelkauskas
© Garland Science
Microbiology: A Clinical Approach
© Garland Science
INDUSTRY AND APPLICATIONS

Industry is using products coming from
biotechnology.

In environmental science:

Renewable energy

Bio
-
based fuels

Bioremediation