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Chapter 12
DNA is the molecule that
carries all of the inherited
information in the cell.
DNA was discovered as
nucleic acidan acidic
material in the nucleus in
the later 1800s.
Its importance was
discovered until later. For
a long time, DNA was
considered too simple to
carry genetic information.
An Experiment
More Experiment
Structure of DNA
DNA is a macromolecule, a
large molecule composed of
many subunits. The subunits
of DNA are nucleotides.
Each nucleotide is composed
of 3 parts: a nitrogenous base,
a sugar (called deoxyribose),
and a phosphate group (which
is a phosphorus atom bonded
to 4 oxygen atoms).
There are 4 kinds of
nitrogenous bases in DNA:
adenine (A), guanine (G),
cytosine (C), and thymine (T).
DNA Replication
Gene Expression
Chapter 13
Each gene is a short section of a
chromosomes DNA that codes for a
polypeptide.
Recall that polypeptides are linear chains of
amino acids, and that proteins are
composed of one or more polypeptides,
sometimes with additional small molecules
attached. The proteins then act as enzymes
or structures to do the work of the cell.
All cells have the same genes. What makes
one type of cell different from another is
which genes are expressed or not
expressed in the cell. For example, the
genes for hemoglobin are on in red blood
cells, but off in muscle and nerve cells.
Expressed = making the protein product.
Genes are expressed by first making an
RNA copy of the gene (transcription) and
then using the information on the the RNA
copy to make a protein (translation).
This process: DNA transcribed into RNA,
then RNA translated into protein, is called
the Central Dogma of Molecular Biology.
RNA
Transcription
More Transcription
RNA processing
Genetic Code
Transfer RNA
Translation
More Translation
Translation, part 3
The ribosome then
slides down the
messenger RNA 1
codon (3 bases).
The first transfer RNA
is pushed off, and the
second transfer RNA,
with 2 attached amino
acids, moves to the
first position on the
ribosome.
Translation, part 4
The elongation cycle
repeats as the
ribosome moves
down the messenger
RNA, translating it
one codon and one
amino acid at a time.
Repeat until a STOP
codon is reached.
Translation, end
Summary of translation
Post-translation
The new polypeptide is now floating loose in the
cytoplasm. It might also be inserted into a
membrane, if the ribosome it was translated on
was attached to the rough endoplasmic
reticulum.
Polypeptides fold spontaneously into their active
configuration, and they spontaneously join with
other polypeptides to form the final proteins.
Sometimes other molecules are also attached to
the polypeptides: sugars, lipids, phosphates, etc.
All of these have special purposes for protein
function.
Mutation
Any change in the base sequence of a DNA molecule is a mutation. Mutation is a completely random process:
any DNA base can be mutated, whether it is in a gene or not.
Basic types:
1. base substitutions: convert one base into another, such as changing an A into a G.
2. Insertions or deletions of large pieces of DNA.
3. Combining parts of 2 different genes together.
Mutations are very common: every cell contains multiple mutations. Also, everyone is genetically different from
every other person due to the accumulation of mutations.
Genetic load: on average, each person has 3 recessive lethal mutations in all cells. We survive because the
dominant normal alleles cover up the recessive lethals. Inbreedingmating with close blood relativesoften
causes defective children because the recessive lethals inherited from the common ancestor become
homozygous.
Many mutations occur in regions where they have no effect: between the genes, or in the introns that are spliced
out of the messenger RNA. Only mutations within genes can affect the organism.
Base substitution mutations within a gene can alter or destroy the genes protein product. The protein may not
function at all, or it might be less efficient, or it might have an altered pH optimum or temperature optimum. Many
of these changes have little or no effect on the organism: these are called neutral mutations, because they are
neither good nor bad for the organism.
The larger changes that occur with insertions, deletions, and rearrangements are usually harmful, because they
usually destroy at least one gene. However, new useful genes sometimes arise from these rearrangements. One
event in particular: attaching the control regions of one gene onto the protein-coding part of another gene. This
causes the protein to be synthesized in a new time and place within the organism.
Rate: for typical genes, base substitutions occur about once in every
10,000 to 1,000,000 cells. Since we have about 6 billion bases of
DNA in each cell, this implies that virtually every cell in your body
contains several mutations. Clearly, most mutations are neutral:
have no effect.
Only mutations in the germ line cells: cells that become sperm or
eggsare passed on to future generations. Mutations in other body
cells only cause trouble when they cause cancer or related
diseases.
Causes: The natural replication of DNA produces occasional errors.
DNA polymerase has an editing mechanism that decreases the rate,
but it still exists.
Radiation and certain chemical compounds also cause mutations.
Chemicals that cause cancercarcinogensalmost all work by
causing mutations.
Gene Control
Chapter 14
Most regulation of genes works by controlling
transcription, the process of making an RNA
copy of a single gene. Thus, a gene is on when
it is being transcribed, and off when it is not
being transcribed.
There are many ways to regulate genesa lot of
contemporary biology research is devoted to
discovering these mechanisms.
I will describe a few simple mechanisms.
Lac Operon
The common gut bacterium
Escherichia coli (E. coli) has
been studied by scientists for a
long time and much is known
about it.
E. coli, like most organisms, used
glucose as its primary food.
However, in the absence of
glucose, it can used lactose (milk
sugar).
Lactose is a disaccharide. E. coli
cells produce an enzyme that
breaks it down into glucose. This
enzyme is called betagalactosidase, and it is made by
a gene called the lac operon.
The lac operon also makes other
proteins that help in the process.
Lac Regulation
Positive Control
X Chromosome Inactivation
More X Inactivation
Hormone Signaling
Genetic Engineering
Chapter 15.
We have been modifying living tings for a long
time: domestication of various plants and
animals, hybridization of different species (such
as horse x donkey = mule), selective breeding for
useful traits.
Recently it has become possible to directly
modify the DNA of living organisms, in the hope
of producing a more useful plant or animal. Also,
we can artificially produce natural body proteins
that can be used as medicinal drugs.
Molecular Cloning
Plasmid Vectors
Restriction Enzymes
DNA Ligase
One major use of cloned genes is to produce large amounts of their protein products
to be used as medicinal drugs.
As an example, human growth hormone is made in the pituitary gland, a very small
organ at the base of the brain. Some people do not produce enough of it, resulting in
very short stature and various health-related issues. HGH injections during childhood
help. However, pituitary glands must be harvested from human cadavers, and are
often contaminated with viruses.
Another example: insulin is needed by people with diabetes. In former times it was
isolated from pigs or sheep. However, the animal forms had a few amino acid
differences from human insulin, and sometimes caused bad immune responses.
The solution to these problems is to isolate the human genes using the techniques of
recombinant DNA, then cause these genes to express themselves, to produce their
protein products. The proteins can then be isolated from the bacteria.
The proteins are the normal human forms, despite having been made in bacteria.
They are not contaminated with human viruses, and they wont cause an immune
reaction.
Getting the genes to express is simple (in principle): RNA copies of genes are made if
they have a promoter sequence in front of them. Similarly, proteins are made if the
messenger RNA has the appropriate signals on it. So, it is merely a matter of
including the proper signal sequences on the plasmid.
But of course: complications arise.
Gene Therapy
Nuclear Cloning