THE RAW MATERIALS OF
BIOTECHNOLOGY
Learning Outcomes
Identify the levels of biological organization and
explain their relationships
Describe cell structure and its significance in
biotechnology research and product development
Discuss the types of organisms researched and the
types of cells grown and studied in biotechnology
facilities plus the product with which they are
associated
Learning Outcomes
Distinguish between the cellular organization of
prokaryotic and eukaryotic cells
List the 4 main classes of macromolecules and
describe their structure and function
Define genetic engineering and identify products
created with this technology
Explain the Central
D
ogma of Biology and its
importance in genetic engineering
Vocabulary
Fluorometer
Organism
Cell
Escherichia coli
Multicellular
Cytology
Anatomy
Physiology
Respiration
Unicellular
Chlorophyll
Photosynthesis
Steroids
nucleotide
Tissue
Organ
Protein
Eukaryote
Protist
Organelles
Mitochondria
Sugar
Starch
Nucleic acid
Pancreas
Hormone
R group
ribonuicleic
acid
Vocabulary
Chloroplast
Cytoplasm
Lysosome
Ribosome
Cell wall
Cellulose
Plasma membrane
Glucose
ATP
Nucleus
Chromosomes
Enzyme
Cellular respiration
Deoxyribose
Hydrophobic
Triglycerides
Ribose
Phospholipids
Pigments
mRNA
Amino acid
Polypeptide
Chinese hamster ovary cells
Vero cells
Prokaryote
Organic
Carbohydrates
Cytoskeleton
Monomer
Polymer
Monosaccharide
Disaccharide
Polysacccharide
Fructose
Sucrose
Lactose
Hydrophillic
Organisms & Their Components
To manufacture biotechnology products for medical,
industrial, or agricultural applications,
biotechnicians
must work either directly or indirectly with
organisms or their components
Entire
multicellular
organisms
Bacteria
Tissues/cells
Organisms & Their
Cmponents
Working in any area of biotechnology requires a
thorough understanding of the characteristics of life
and the structures that compose organisms
You must have at least a minimal understanding of
Biochemistry: organic molecules structure, function, &
interactions
Cytology: branch of life science, which deals with the
study of cells in terms of structure, function and
chemistry
Anatomy: structure of living things and their components
Physiology: the processes and functions of living systems
The Living Condition
Biologists estimate that there are well in excess of
20million different species, with some estimates
being as high as 150 million
All living things can be classified into one of the
following categories
Prokaryotes
Eukaryotes
Eubacteria
Archaeabacteria
Protists
Plants
Animals
Fungi
The Living Condition
Biologists estimate that there are well in excess of
20million different species, with some estimates
being as high as 150 million
All living things can be classified into one of the
following categories
Prokaryotes
(Unicellular)
Eukaryotes
(mostly
multicellular
)
Eubacteria
Archaeabacteria
Protists
Plants__
Animals__
Fungi
Characteristics of Life
Growth & Development
Reproduction
Organized structure composed of 1 or more cells
Response to stimuli
Conversion of energy
Respiration
Metabolism
Catabolism
Anabolism
Here We Go Again! Cells & Their Parts
The cells of both unicellular and
multicellular
organisms
are tiny microscopic factories that produce thousands of
different molecules
Biotechnology companies exploit the
biologiical
manufacturing capabilities of cells and trick them into
producing particular molecules in large quantities
These become biotechnology products
Organelles: a specialized subunit within a cell that has
a specific function, and is usually separately enclosed
within its own lipid
bilayer
The name
organelle
comes from the idea that these
structures are to cells what an organ is to the body
Organelles involved in Energy
Production
A membrane bound organelle
consisting of an outer
membrane and a highly
folded inner
-
membrane; it is
responsible for generating
large amounts of ATP through
the process of aerobic cellular
respiration
Found in all eukaryotic cells
ATP: adenosine
triphosphate
a nucleotide that serves as
the main source of usable
energy for ALL living things
A specialized organelle
in plants and some
protists
that uses the
energy from sun light to
convert
inorgaic
molecules (CO
2
& H
2
O)
into energy storing
organic molecules
(glucose) through the
process of
photosynthesis
Mitochondria
Chloroplast
Nucleus: The Organelle that Stores the
Hereditary Material
a double lipid
bilayer
that encloses
the genetic material in eukaryotic
cells.
serves as the physical barrier,
separating the contents of the nucleus
from the
cytosol
nuclear pores are inserted in the
nuclear envelope, which facilitate
and regulate the exchange of
materials (proteins such as
transcription factors, and RNA)
between the nucleus and the
cytoplasm.
The outer membrane is continuous
with the rough endoplasmic reticulum
The outer and inner nuclear
membrane are fused at the site of
nuclear pore complexes.
DNA: a nucleic acid that contains
the genetic instructions used in the
development and functioning of
all known living organisms
main role is the long
-
term
storage of information
Used as a template to make RNA
through the
priocess
of
transcription
Nucleolus: a non
-
membrane
bound structure
composed of
proteins and nucleic acids found
within the nucleus.
Ribosomal RNA (
rRNA
) is
transcribed and assembled within
the nucleolus, forming
ribosomes
Nuclear Envelope
Nuclear Contents
Ribosomes
: involved in Protein Synthesis
Non
-
membrane bound organelles composed of
rRNA
& protein that provide the appropriate
microenvironment to catalyze peptide bond
formation between amino acids based on the
instructions it reads on an mRNA molecule through
the process of translation
Chemistry Connect: Remember, all organic polymers are
formed through condensation reactions, aka
dehydration synthesis
Organelles involved in Trafficking
A highly folded lipid
bilayer
whose surface
ois
studded with
protein
-
manufacturing
ribosomes
giving it a "rough" appearance
Ribosomes
bound to the RER at
any one time are not a stable
part of this organelle's structure
as
ribosomes
are constantly
being bound and released from
the membrane.
A ribosome only binds to the ER
once it begins to synthesize a
protein destined for the
secretory
pathway
The Golgi apparatus
processes and packages
macromolecules, such as
proteins and lipids, after
their synthesis and before
they make their way to
their destination; it is
particularly important in
the processing of proteins
for secretion.
Rough Endoplasmic Reticulum
Golgi Apparatus
Organelles involved in Metabolism
Functions in several metabolic
processes, including synthesis
of lipids and steroids,
metabolism of carbohydrates,
regulation of calcium
concentration, drug
detoxification, attachment of
receptors on cell membrane
proteins, and steroid
metabolism.
It is connected to the nuclear
envelope.
Lysosomes: contain acid enzymes
to break up waste materials and
cellular debris.
found in animal cells, while in
yeast and plants the same roles
are performed by lytic
vacuoles
.
digest excess or worn
-
out
organelles, food particles, and
engulfed viruses or bacteria
The membrane around a
lysosome allows the digestive
enzymes to work at the 4.5 pH
they require.
Smooth Endoplasmic Reticulum
Lysosome
Metabolism: is the set of chemical reactions that
happen in living organisms to maintain life
is the set of pathways
that break down
molecules into smaller
units
These reactions
release energy
the set of metabolic
pathways that
construct molecules
from smaller units.
These reactions require
energy
Catabolism
Anabolism
Critical Thinking: Which
organelles are catabolic and
which are anabolic?
The
Endomembrane
System
Is composed of the different membranes that are
suspended in the cytoplasm within a eukaryotic cell.
These membranes divide the cell into functional and
structural compartments, or organelles.
In eukaryotes the organelles of the
endomembrane
system include: the nuclear envelope, the
endoplasmic reticulum, the Golgi apparatus,
lysosomes
, vacuoles (plants), vesicles (all
eukaryotes), and the cell membrane
Cellular Membranes
Acts as a semi
-
permeable barrier that
regulates what enters or leaves the cell or
organelle
They consist of phospholipids which
spontaneously arrange so that the hydrophobic
"tail" regions are shielded from the surrounding
polar fluid, causing the more hydrophilic
"head" regions to associate with the
cytosolic
and extracellular faces of the resulting
bilayer
.
Things in/on Cellular Membrane
Proteins: function in cellular communication, catalyzing
reactions, cell, signaling, cell movement
4 main types
Receptor proteins bind signal molecules and initiate signal
transduction pathways
Enzymes: speed up chemical reactions
Channel proteins: open or close to let target molecules in or out in
response to a signal
Marker proteins: specific for each cell type; also, help the immune
system distinguish self from non
-
self
Cholesterol: establishes proper membrane permeability
& fluidity
Cell Walls
All living things except animals and some
protists
have
cell walls
Cell walls are
semipermeable
boundaries that are more
rigid than, and surround the plasma membrane
provides structural support and protection, and also acts as
a filtering mechanism
Keeps cells from bursting in hypotonic environments
Different organisms have cell walls made of different
organic polymers
Plants: cellulose
Bacteria:
peptidoglycan
Fungi (some): chitin
Cell
Tissue
Organ
Organism
Depending on the type of cell hundreds of different
molecules are being
manuifactured
at any given
moment
Some molecules are unique to a specific type of cell,
others are produced in all cells
Example: Pancreas
organ of the digestive system that
lies in your abdomen, behind your
stomach,with
2 main
functions:
producing digestive enzymes to break down food; and
producing the hormones insulin and glucagon to control
sugar levels in your body.
Case in Point: Pancreas
Cells
Liver cells are called
hepatocytes
α
-
cells: secrete glucagon
β
-
cells secrete insulin
Acinar
cells: secrete enzymes that
allow your body to digest protein, fat
and starch from your food
Tissue
Islets of
langerhans
Secrete hormones
Acinar
tissue
Secrete digestive enzymes
Organ
Pancreas
Organism
People
A closer look at the BIG Picture
When you, the organism, eats a meal, your stomach
(with the help of digestive enzymes from your
pancreas and other organs) break down the
complex carbohydrates into glucose
Glucose is absorbed through the intestines (small I
think) into the blood stream
Rising glucose in the blood stream is detected by
the pancreas as the glucose binds glucose receptors
on pancreatic cells. This stimulates
β
-
cells
to
release insulin
Insulin:
pancreatic homeostasis
Insulin works by improving the uptake of glucose from
the blood across cell membranes and into the cells of
the body by binding to insulin receptors on cells
Specifically, when insulin binds the insulin receptor, the
receptor protein transports phosphate groups from ATP
to other proteins in the cell, initiating signal transduction
pathways that open glucose channels allowing passive
diffusion of glucose into the cell
Once in the cells, the glucose is used as the energy to fuel
the cells doing their different jobs or is stored in the liver or
muscle cells as glycogen.
This results in the glucose level of the blood dropping, which
then triggers the pancreas to switch off the release of insulin
Critical Thinking
Insulin and glucagon are both hormones that work
together to regulate blood glucose levels. Knowing
how insulin works, and that glucagon's effect is
opposite that of insulin, propose a mechanism by
which glucagon may work
Hint: remember that unused glucose is converted to and
stored in the form of glycogen in the pancreas
Glycogenolysis
Glucagon helps maintain the level of glucose in the
blood.
Glucose is stored in the liver in the form of
glycogen, which is a starch
-
like polymer chain made
up of glucose molecules.
Liver cells (
hepatocytes
) have glucagon receptors.
When glucagon binds to the glucagon receptors,
the liver cells convert the glycogen polymer into
individual glucose molecules, and release them into
the bloodstream, in a process known as
glycogenolysis
.
Why all this matters
The purpose of biotechnology is to increase quality
and/ or quantity of life
Life is dependant on the bodies abilities to
Maintain homeostasis
Transform energy
Reproduce
Grow and develop
Respond to stimuli
Internal: for example a change in levels of blood sugar
External: for example a change in temperature
Biotechnology focuses on
making organisms better able
to accomplish one or more of
these tasks
Central Dogma
If the goal of biotechnology is to increase an
organisms ability to accomplish one or more of the
characteristics of life, then it is important to
understand how the organism naturally carries these
tasks out!
One of the most important aspects of understanding
this is to understand the central dogma of biology
Chromosomes & Genes
We know chromosomes are in the nucleus
We know genes are on chromosomes, and are
segments of DNA that code for protein
What we need to look at more specifically is HOW
the DNA instructs your body to build a protein
Central Dogma: an Overview
DNA is made up of 2 strands of nucleotides that coil
around each other forming a double helix
Each strand is composed of varying arrangements
of 4 nucleotides
Adenine
Cytosine
Thymine
Guanine
When genes are used to instruct the body to build a
protein, they must be
transcribed
into a
complementary sequence of
messenger RNA
The mRNA transcript is made of nucleotides
Adenine
Cytosine
Uracil
Guanine
Central Dogma: an Overview
This mRNA can leave the nucleus and be read by a
ribosome
which actually constructs the protein
through the process of
translation
In translation, the ribosome links amino acids
together in the order specified by the sequence of
nucleotides in the mRNA
These amino acids are held together by special
covalent bonds called
peptide bonds
There are
20 different amino a
cids
A chain of amino acids are called a
polypeptide
Nucleus
Ribosome in cytoplasm or on RER
Central Dogma
The process of Gene Expression is universally found
in all cells
How the DNA code is rewritten into mRNA and then
decoded into a protein
It is called “The Central Dogma of Biology” because
it helps explain how virtually all molecules are
made either
directly protein & nucleic acids
or indirectly carbohydrates and lipids because their
synthesis is controlled by the proteins
Tying it together
DNA is used to make protein
Proteins do most of the work of the cell
The other organic molecules are important too, but
because their production/ destruction is controlled by
proteins we focus on the protein
The cell has specialized organelles, containing
different collections of proteins, giving each
organelle a unique function based on the proteins
(and other organic molecules) in/on it
The organelles work together to help the cell carry
out all of the characteristics of life
Tying it together
Not all cells, even within the same individual, are
the same
Some cells become specialized to perform a
specific function
β
-
cells:
Secrete insulin
Rods: sense light
Cells are able to perform these specialized
functions BECAUSE of the proteins they produce
Different types of cells produce different
types/quantities of proteins at different times
Critical Thinking
If all cells within one organism have the same set of
DNA (in other words, the DNA in your nerve cells is
identical to the DNA in the epithelium of your little
toe), how is it that all cells are not the same?
Cells Commonly Used in Biotechnology
In biotech applications, some cells are used more than
others
Chinese Hamster Ovary (CHO)cells
Vero Cells: African green monkey kidney epithelial cells
HeLa
cells: human epithelial cells
Fungal cells
Aspergillus
yeast
Prokaryotic cells
E. coli
Staphylococcus
Streptococcus
In Other Words
You need to know how
to keep cells alive in
culture, in order to do
most biotechnology
research
NOTE
On page 46 of your text book it claims that
“Due to their lack of mitochondria, bacteria conduct
ONLY anaerobic respiration”
This is WRONG & the author is a MORON
There are SOME strictly anaerobic bacteria but
MOST need or at least can survive in oxygen
environments!
Read section 2.3 on your own!
Things to pay attention to
Carbohydrates
Glycogen: what is it
Structural polysaccharides of cell wall
How polysaccharides
interefere
with purification procedures
The text book reads “Cells break the bonds in glucose, releasing energy in a
form that cells can use” why do I hate that definition
Figure 2.23
lipids
why are lipids referred to as hydrocarbons
The 3 general groups of lipids and their functions
Triglycerides
Phospholipids
Steroids
Structure of a
phospholipid
“DNA is the Flash, Proteins are the Cash!”
Water makes up 75% of a cells mass
Of 25% of dry mass, 75% of it is protein
In biotechnology, proteins are often the
manufactured product
Often, 50
-
75% of a companies staff is devoted to
protein research (proteomics)
Proteins fall into 9 different categories
depending on their function
Structural
:
proteins with the primary purpose of
producing the essential structural components of the cell
Enzyme
:
speed up chemical reactions by lowering activation energy
Transport
:
involved in the movement of substances across
biological membranes; or carries specific molecules (example
oxygen is transported by hemoglobin)
Contractile
:
involved in muscle contraction, also in the functioning
of the mitotic spindle
Hormone
:
a chemical released by a cell or a gland in one part
of the body that sends out messages to affect cells in other parts of
the organism
Proteins fall into 9 different categories
depending on their function
Antibody
:
used by the immune system to identify and
neutralize foreign objects, such as bacteria and viruses
Pigment
:
a material that changes the color of reflected or
transmitted light as the result of wavelength
-
selective absorption
Marker/ Recognition
:
proteins usually found on the
extracellular membrane that allows other molecules to identify the
cell type
Toxins:
small molecules, peptides, or proteins that are capable of
causing disease on contact with or absorption by body tissues
interacting with biological macromolecules such as enzymes or
cellular receptors
Proteins
Grouping is based on functions, and often proteins
within the same group are more similar to each other
than to proteins in different groups
Note: some proteins can have different functions, even
within the same organism, depending on which cell they
are produced in
A typical cell produces about 2,000 different proteins
Proteins are the work horses of the cell, each able to
accomplish its task due to its specific structure and
properties
Proteins
The structure of a protein is determined by the amino
acid sequence
Chains of amino acids are called polypeptides
Polypeptides are not functional until they fold into their
3
-
dimensional shape
Folded polypeptide chains are called proteins
The way a polypeptide folds into a functional protein is
determined by the sequence of the amino acids and
their properties
The sequence of amino acids is determined by the
sequence of mRNA which is determined by the
sequence of DNA
There are 20
different amino
acids
All amino acids
have the same
basic structure
R
-
Groups
R
-
groups are the chemical side groups on an amino
acid that varies between different amino acids
It is the R
-
group that is responsible for the unique
characteristics of each amino acid
The chemical nature of each R
-
group results in
attractions and repulsions between certain amino acids
Example: the negatively charged R
-
group of
glutamic
acid
is attracted to the positively charged R
-
group of
arginine
The various folding patterns of each protein are a result of
these interactions
R
-
Groups
The R
-
groups of protein chains can interact between
proteins as well
Many proteins function by attracting or repelling
other protein chains
Many recognition proteins, antibodies, enzymes, and
protein hormones function because the R
-
group on
one of their amino acids is strategically placed
(active site) to interact with its target molecule
(
ligand
)
Nucleic Acids
Information carrying molecules that direct the
synthesis of all cellular molecules, including
themselves
Ultimately, each protein, carbohydrate, and lipid
molecules production can be traced back to genetic
information stored in the sequence of DNA, which is
packaged into chromatin or chromosomes,
depending on what stage of the cell cycle the cell is
in
Quick Comparison of Prokaryotic Chromosomes
to Eukaryotic Chromosomes
Chromosome located in the
cytoplasm in a region known
as the
nucleoid
1 chromosome (may have
additional plasmids)
Chromosome is circular
No introns, exons only
DNA is NOT associated with
histone
proteins
Chromosomes found in the
nucleus
# of chromosomes varies
with species
Chromosomes are linear
DNA contains both introns &
exons
DNA is associated with
histone
proteins
-
Genetic
information
-
Contains
A,C,T,G
-
Composed
of
nucleotides
Prokaryotes
Eukaryotes
Overview of Gene Expression
DNA is composed of long stretches of nucleotides
covalently linked together
Genetic information lies in the sequence of
nitrogenous bases
There are 5 nitrogenous bases
DNA: adenine, cytosine, guanine, thymine
RNA: adenine, cytosine, guanine,
uracil
Cytosine on 1 strand always pairs with guanine on the other
strand
Adenine pairs with either thymine (if you’re copying DNA) or
Uracil
, (if your transcribing RNA
DNA is the template to build RNA
A
A
T
C
C
G
T
C
T
A
T
G
T
T
A
G
G
C
A
G
A
T
A
C
REPLICATION
A
A
U
C
C
G
U
C
U
A
U
G
TRANSCRIPTION
TRANSLATION
The sequence of bases is read in groups
of 3
Each group of 3 bases is called a
codon
and specifies 1 amino acid
Asparagine
Proline
Serine
Methionine
Occurs on a ribosome in the
cytoplasm or attached to the
RER
Occurs in the nucleus
Peptide bonds are a type of covalent bond that hold
amino acids together
What type of reaction forms peptide bonds?
Genetic Code
This is how a molecule with only 4 nitrogenous bases
can code for over 25,000 different proteins!
Because RNA and so the gene is “read” in groups of
3
We can use this knowledge to perhaps not just treat
but actually fix genetic mistakes and or give
organisms new characteristics that make them more
fit, substantially improving the quality and quantity
of life
Genetic Engineering
Companies employ genetic engineers to isolate and
alter the DNA codes for a particular protein or
group of proteins
Sometimes the protein, like insulin, is the product
itself
The goal, then, is to produce it in sufficient quantities
to sell in the marketplace and make a profit
The “New” Biotechnology
Organisms & their products have been harvested
and improved for centuries
The most significant breakthrough in the
manipulation of plant and animal cells occurred
when scientists learned how to move pieces of DNA
within and between organisms
The key to this discovery was restriction enzymes
and DNA
ligase
, which allow us to cut and paste
DNA to make
rDNA
Recombinant DNA
rDNA
usually contains fragments from different
organisms
They are usually novel molecules not seen in
existence anywhere else, and thus are referred to
as being “engineered”
DNA fragments containing genes of interest can be
pasted into vectors and carried back into cells
Once in cells they are transcribed and translated
into protein molecules that the recipient cell has
never produced
This organism has been “genetically engineered”
The 1
st
Genetic Engineering Experiment
Occurred in 1973 when 3 scientists
Stanley Cohen (Stanford)
Herb Boyer (University of California)
Paul Berg (Stanford)
Excised (cut out) a segment of amphibian DNA from
the African Clawed Toad,
Xenopus
, and pasted it
into a small ring of bacterial DNA called a plasmid
The new recombinant plasmid (r
-
plasmid) was
placed in an
E. coli
cell which then transcribed the
DNA into ribosomal RNA
Human Insulin
The 1
st
genitically
engineered product to reach the
market place was human insulin
Scientists used similar methods as before to excise a
healthy human insulin gene using restriction enzymes,
paste it into a bacterial plasmid using DNA
ligase
, and
then transferred the plasmid to E. coli cells which then
transcribed the DNA into RNA, and then translated the
mRNA into the insulin protein
The cells were grown in large volumes, and then the
insulin was purified out of the cell culture
The FDA approved r
-
human insulin (
rhinsulin
)in 1982
Genentech
In 1976 Robert Swanson & Herb Boyer founded the 1
st
biotechnology company called Genentech in San
Francisco
Their 1
st
product was
rhInsulin
Genentech has grown into one of the largest
pharmaceutical companies in the world
It currently markets or is developing
t
-
PA: treatment of
heartattachs
Nutropin
: a human growth hormone
Rituxin
: an antibody that targets cancer
cellsin
B
-
cell non
-
Hodgkins
lymphoma
Pulmozyme
: for cystic fibrosis treatment
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