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DNA

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

How it was learned that DNA was the hereditary


material: experiments by Griffith and then by Avery,
Macleod, and McCarthy in the 1920s through
1940s.
They used bacteria called Streptococcus
pneumoniae, one cause of pneumonia. They had 2
strains: R (formed rough colonies) and S (formed
smooth colonies due to a polysaccharide coat).
When injected into mice, S bacteria caused
pneumonia and killed them. R bacteria didnt hurt
the mice.
When he killed the S bacteria by heating them, they
no longer caused pneumonia. Same for R bacteria.
Heres the important result: if he injected live R
along with the heat-killed S bacteria, the mice
developed pneumonia and died. And, they
contained live S bacteria.
What happened: the hereditary material from the S
bacteria survived the death of the bacteria
themselves, and it transformed the live R bacteria
into S bacteria.. Demonstrated that the hereditary
material is separate from the property of being
alive.

More Experiment

Later work showed that the hereditary


material was DNA.
Crude extracts were made from the S cells:
breaking them apart. The extracts
transformed live R cells into S cells and
killed the mice, just like heat-killed S cells.
The extracts were treated with various
enzymes known to digest different cellular
components: protein, RNA, DNA, etc.
DNAase, the enzyme that digested DNA,
stopped the transformation effect, but none
of the other enzymes did. This
demonstrated that DNA was the active
material in transformation, the hereditary
material.
Numerous other experiments, using
different organisms and procedures,
continued to show that DNA, and not protein
or some other type of molecule, was
responsible for inheritance.

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).

More DNA Structure

The nucleotides are joined into long


chains that connect the phosphate of
one nucleotide to the sugar of the next
nucleotide.
The nitrogenous bases of 2 different
chains pair with each other, giving a
DNA molecule that has 2 sugarphosphate chains on the outside, with
bases paired in the center.
Base pairing occurs by hydrogen
bonds: partial positive and negative
charges attract each other. Hydrogen
bonds are weak, but there are lots of
them in a DNA molecule.
The 2 chains are anti-parallelthey
run in opposite directions. They are
twisted together into a corkscrew
shape: a double helix.
Base pairing is very specific: A pairs
with T, and G pairs with C.

DNA Replication

How DNA makes copies of itself.


Occurs during the S phase of the cell cycle,
when each chromosome starts with 1
chromatid and ends with 2 identical
chromatids. Each chromatid is 1 molecule
of DNA.
Involves an enzyme: DNA polymerase.
The DNA double helix unwinds into 2
separate strands, and a new strand is build
on each old one. Thus, each new DNA
molecule consists of 1 old strand plus 1 new
strand. This is called semi-conservative
replication.
DNA polymerase makes the new strands,
using the old strands as a template, with
normal base pairing: A with T, and G with C.
The energy for this comes from the
nucleotide precursors. They all have 3
phosphates on them, like ATP, and 2 of the
phosphates are removed to make the DNA.

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

RNA is a nucleic acid, like DNA, with a few small differences:


RNA is single stranded, not double stranded like DNA
RNA is short, only 1 gene long, where DNA is very long and
contains many genes
RNA uses the sugar ribose instead of deoxyribose in DNA
RNA uses the base uracil (U) instead of thymine (T) in DNA.
There are 3 main types of RNA in the cell:
1. messenger RNA: copies of the individual genes
2. ribosomal RNA: part of the ribosome, the machine that
translates messenger RNA into protein.
3. transfer RNA, which is an adapter between the messenger
RNA and the amino acids it codes for.

Transcription

Transcription is the process of making an


RNA copy of a single DNA gene.
The copying is done by an enzyme: RNA
polymerase. Recall that in replication, a
DNA copy of DNA is made by the enzyme
DNA polymerase.
The bases of RNA pair with the bases of
DNA: A with T (or U in RNA), and G with C.
The RNA copy of a gene is just a
complementary copy of the DNA strand.
RNA polymerase attaches to a signal at the
beginning of the gene, the promoter. Then
RNA polymerase moves down the gene,
adding new bases to the RNA copy, until it
reaches a termination signal at the end of
the gene.

More Transcription

RNA processing

Oddly, most genes in eukaryotes are not


continuous. They are interrupted by long regions of
DNA that dont code for protein, called introns.
Introns have no known function. The useful parts
of the gene, the parts that code for proteins, are
called exons. Some genes are more than 99%
introns, with only 1% of the gene useful: the cystic
fibrosis gene is like this.
The entire gene, introns and exons, is transcribed
into an RNA copy, but the introns need to be
removed before it can be converted to protein.
After transcription, snips out the introns, leaving
only the protein coding portion of the gene in the
RNA.
Also, the cell adds a protective cap to one end, and
a tail of As to the other end. These both function to
protect the RNA from enzymes that would degrade
it starting on an end and moving inward.
Thus, an RNA copy of a gene is converted into
messenger RNA by doing 2 things: 1. cut out the
introns. 2. add protective bases to the ends.
Transcription of RNA processing occur in the
nucleus. After this, the messenger RNA moves to
the cytoplasm for translation.

Genetic Code

There are only 4 bases in DNA and


RNA, but there are 20 different amino
acids that go into proteins. How can
DNA code for the amino acid
sequence of a protein?
Each amino acid is coded for by a
group of 3 bases, a codon. 3 bases of
DNA or RNA = 1 codon.
Since there are 4 bases and 3
positions in each codon, there are 4 x
4 x 4 = 64 possible codons.
This is far more than is necessary, so
most amino acids use more than 1
codon.
3 of the 64 codons are used as STOP
signals; they are found at the end of
every gene and mark the end of the
protein.
One codon is used as a START signal:
it is at the start of every protein.

Transfer RNA

Transfer RNA molecules act as adapters


between the codons on messenger RNA and
the amino acids. Transfer RNA is the physical
manifestation of the genetic code.
Each transfer RNA molecule is twisted into a
knot that has 2 ends.
At one end is the anticodon, 3 RNA bases that
matches the 3 bases of the codon. This is the
end that attaches to messenger RNA.
At the other end is an attachment site for the
proper amino acid.
A special group of enzymes pairs up the proper
transfer RNA molecules with their corresponding
amino acids.
Transfer RNA brings the amino acids to the
ribosomes, which are RNA/protein hybrids that
move along the messenger RNA, translating the
codons into the amino acid sequence of the
polypeptide.

Translation

Three main players here:


messenger RNA, the
ribosome, and the transfer
RNAs with attached amino
acids.
First step: initiation. The
messenger RNA binds to a
ribosome, and the transfer
RNA corresponding to the
START codon binds to this
complex. Ribosomes are
composed of 2 subunits (large
and small), which come
together when the messenger
RNA attaches during the
initiation process.

More Translation

Step 2 is elongation: the ribosome


moves down the messenger RNA,
adding new amino acids to the growing
polypeptide chain.
The ribosome has 2 sites for binding
transfer RNA. The first RNA with its
attached amino acid binds to the first
site, and then the transfer RNA
corresponding to the second codon
bind to the second site.
The ribosome then removes the amino
acid from the first transfer RNA and
attaches it to the second amino acid.
At this point, the first transfer RNA is
empty: no attached amino acid, and
the second transfer RNA has a chain
of 2 amino acids attached to it.

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

The final step in translation is


termination. When the
ribosome reaches a STOP
codon, there is no
corresponding transfer RNA.
Instead, a small protein called
a release factor attaches to
the stop codon.
The release factor causes the
whole complex to fall apart:
messenger RNA, the two
ribosome subunits, the new
polypeptide.
The messenger RNA can be
translated many times, to
produce many protein copies.

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.

Mutation Causes and Rate

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 Operon Basics

The operon itself consists of 3


regions that code for protein, called
Z, Y, and A. The Z gene codes for
the important enzyme, betagalactosidase.
A single messenger RNA is made
from the entire lac operon.
Transcription of the RNA starts at the
promoter at the left end of the gene.
The operator is a region of DNA
between the promoter and the Z, Y,
and Z genes. It is an important part
of the regulatory system.
Another gene makes the lac
repressor, the protein involved in
gene regulation.

Lac Regulation

To conserve its resources, the E. coli


need to have the lac operon ON when
lactose is present and OFF when
lactose is absent.
To accomplish this, the repressor
protein can be in two different states:
the repressor can either bind to
lactose, or it can bind to the operator
DNA sequence.
When lactose is present, the repressor
binds to lactose and not to the
operator. This allows RNA polymerase
to transcribe the gene: it is ON.
When lactose is absent, the repressor
binds to the operator. This blocks RNA
polymerase from transcription, and the
gene is OFF.
If lactose is added, the repressor falls
off the operator and binds the lactose,
which allows transcription to start.

Positive Control

The lac operon is an example of negative


regulation: the regulatory protein (the lac repressor)
causes transcription to stop.
Positive control, where the regulatory protein
causes transcription to start, is more common.
The lac operon of E. coli also has an example of
positive control. When the cells glucose level is
high, it doesnt need to use lactose at all. So, only
when the glucose level drops is it necessary to try
using lactose.
The positive control mechanism turns the lac
operon ON when the glucose level drops.
This mechanism uses a different protein, called
CAP, and the signaling molecule cyclic AMP
(cAMP).
cAMP is generated when the glucose level is low.
cAMP then binds to CAP, and the cAMP-CAP
complex attaches itself to the promoter. This
complex attracts RNA polymerase and allows
transcription to occur at a high rate.
The negative regulation system acts on top of the
positive system: transcription is allowed whenever
glucose is low, but only occurs is lactose is present.

X Chromosome Inactivation

Many genes in multicellular organisms are


shut down permanently in different tissues.
They are never needed because they make
products used only in other types of cells.
A model for this permanent inactivation is
the X chromosome in females. Males have
only 1 X and females have 2 Xs. For any
other chromosome, this imbalance would be
lethal, but it is the normal condition of the X.
How can this occur?
In every female body cell, only 1 X is active.
The other X gets converted into a
condensed, inactive blob on the nuclear
membrane called a Barr body. This is a
simple way to telling male cells from female
cells: female cells have Barr bodies and
male cells dont.
Only one X is active in every cell: all others
are converted to Barr bodies. People with
XXY (Klinefelters syndrome) are male in
appearance, but their cells have Barr
bodies. People with Turners syndrome (XO,
only 1 X) are female but have no Barr
bodies. People with 3 Xs (XXX) have 2 Barr
bodies in each cell.

More X Inactivation

When the embryo has about 200 cells, each cell


independently inactivates all but 1 X chromosome. The
inactive X stays inactive in all cells descended from the
original cell, throughout the individuals life.
The inactivation can lead to interesting effects. Tortoiseshell
cats are a mixture of black and orange. They are always
female (occasionally Klinefelters XXY males), because
they need 2 Xs. The gene for coat color is on the X and it
has a black allele and an orange allele. A heterozygote has
one black and one orange allele. But: only 1 X is active in
each cell, which means that either the black allele is active
or the orange allele is active, but not both. So, as the cells
of the embryo develop into patches of skin, the active allele
is expressed and you get a black and orange pattern.
Calico cats also have white spots. Their black and orange
pattern is the result of X inactivation, but the white spots are
due to another, autosomal gene. Calico cats are also
always female.
There is a human condition called anhidrotic ectodermal
dysplasia that cause a loss of sweat glands in the skin. In
females it leads to patches with and without sweat glands. It
is lethal in males.

Hormone Signaling

Hormones are small molecules that


circulate in the blood and alter the
expression of genes in many tissues.
Hormones are either steroids (lipids
with 4 rings of atoms in a characteristic
shape) or peptides (small proteins).
Steroid hormones can enter the cell
directly through the membranes. Once
in the cell, they bind to receptor
proteins, then move to the nucleus
where they stimulate transcription.
Peptide hormones bind to receptor
proteins on the surface of the cell. The
receptor proteins then activate other
proteins within the cell, in a cascade
that ends up activating transcription
factors in the nucleus. Transcription
factors stimulate RNA transcription of
particular genes.

Plant Response to Light

Many plants will only flower when days are


short, while other plants require a long day.
Plants are able to determine the length of
the dark period. Even a very short 10
second pulse of light in the middle of the
dark period prevents short day/long night
plants from flowering.
The mechanism for determining day length
uses a blue-green pigment called
phytochrome. Phytochrome is activated
by red light, which dominates during the
day. The active phytochrome slowly
converts back to the inactive form during the
night. The amount of active phytochrome
left at the end of the night is proportional to
the length of the night.
When present at the appropriate level,
phytochrome stimulates enzymes in the
plant cell to start or stop transcription.
Different plants have different critical levels
of active phytochrome.

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

Molecular cloning means taking a


gene, a piece of DNA, out of the
genome and growing it in bacteria.
The bacteria (usually E. coli) produce
large amounts of this particular gene.
The cloned gene can then be used for
further research, or to produce large
amounts of protein, or (sometimes) to
be inserted into cells that lack the gene
(people with genetic disease, for
example).
The basic tools:
1. plasmid vector: small circle of
DNA that grows inside the bacteria. It
carries the gene being cloned
2. Restriction enzymes: cut the
DNA at specific spots, allowing the
isolation of specific genes.
3. DNA ligase, an enzyme that
attached pieces of DNA together.

Plasmid Vectors

Bacterial chromosomes are large DNA


circles. Bacteria have a single
chromosome.
Plasmids are small circles of DNA inside
bacteria that replicate independently of the
bacterial chromosome. They exist naturally,
and usually confer some useful but not
necessary trait on the bacteria: drug
resistance, for example. Each plasmid only
has a small number of genes on it.
Some plasmids can produce hundreds of
copies of themselves inside each bacterial
cell.
It is possible to insert foreign DNA into the
plasmid. This DNA becomes part of the
enlarged circle of the plasmid. It replicates
along with the plasmid.
The plasmid DNA can be manipulated in
vitro, outside the cell, then put back into the
cell through the process of transformation.
Because bacteria can be grown quickly and
easily, you can produce large quantities of
any DNA inserted into a plasmid

Restriction Enzymes

Restriction enzymes are part of the


defense system of bacteria: they
digest foreign DNA that enters the
bacterial cell.
Each species of bacteria has its own
set of restriction enzymes. Each
enzyme cuts DNA at a specific short
base sequence. For instance, EcoR1
cuts the DNA at the sequence
GAATTC, and BamH1 cuts at
GGATCC. There are hundreds of
restriction enzymes known.
Using properly chosen enzymes, the
gene you want can be cut out of the
chromosome intact, with very little
extra DNA.
Many restriction enzymes give a
staggered cut across the DNA double
helix. This produces short single
stranded regions, called sticky ends.
The ends are sticky because they
spontaneously pair with similar ends.

DNA Ligase

ligate means to tie together.


DNA ligase is an enzyme that
attaches 2 pieces of DNA
together. It forms covalent
bonds between the sugars and
phosphates of the DNA
backbones.
Especially useful: if 2 different
DNAs were cut with the same
restriction enzyme, they will
have matching sticky ends.
DNA ligase can then combine
these 2 different DNA
molecules into a single DNA.
This hybrid DNA molecule is
called recombinant DNA.

The Cloning Process


1. Cut genomic DNA with
a restriction enzyme.
2. Cut plasmid vector with
the same restriction
enzyme.
3. Mix the two DNAs
together and join them
with DNA ligase.
4. Put the recombinant
DNA back into E. coli by
transformation.
5. Grow lots of the E. coli
containing your gene.

Using Cloned DNA

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

One way to cure genetic diseases is to


insert good (non-mutant) copies of the
defective gene into the cells of the
affected person.
Big problem: how to get the gene into
all of the cells.
In mice, inject the gene directly into the
zygote. The gene incorporates into
the chromosomes at some random
location, and (usually) functions.
Humans usually dont know about the
presence of the disease until the baby
is born.
In some cases, blood cells can be
used. Blood cell precursors are in the
bone marrow, which can be extracted,
have recombinant genes inserted, then
put back into the body.

Gene Therapy Problems

Two big problems:


1. The recombinant genes insert
into random locations. Sometimes
they insert into oncogenes, which
causes cancer. Leukemia usually,
cancer of the blood cells.
2. There are very few blood stem
cells in the body. Stem cells divide
repeatedly and never differentiate into
the final blood cells. Genes inserted
into stem cells are permanently in the
body. Genes inserted into cells that
have already started differentiating into
blood cells will stay in the body for a
few weeks, then be lost as the blood
cells wear out and die.
The bottom line (so far): gene therapy
has not been very successful except in
a small handful of cases.

Genetically Modified Plants

It is fairly easy to insert genes into


plants, using a special plasmid vector
derived from crown gall tumors. This
vector grows in bacteria, but transfers
its DNA into the plant genome after
infection.
One use of this techniques is to insert
nutrient genes into plants. For
instance, rice is the staple food for a
large part of the worlds population. It
contains no carotene, the orange
pigment that is the precursor for the
main visual pigment retinol. People
who live on rice alone often develop
blindness because they dont eat
enough carotene.
Golden rice was developed to solve
this problem: genes fro producing
carotene were put into rice. This
pigment gives the rice its color.
Problems: will people eat this oddlycolored food? Will it work well in
cooking? Will they accept
Frankenfood?

More Plant Genetic Engineering

In the US, the main uses of genetically modified


crop plants are herbicide resistance and insect
resistance. If your crop plants are resistant to a
herbicide that kills the weeds, you can spray more
effectively. Similarly, plants that are given genes for
insect resistance dont need to be sprayed with
insecticides.
Very effectivefarmers find these plants cheaper to
grow.
However, there is a lot of resistance to eating these
plants in Europe. Maybe the genes will somehow
leak out and affect peoplewe digest DNA down to
nucleotide subunits before taking it into our bodies,
so this shouldnt happen. Maybe it will affect other
plantsprobably does happen naturally at a slow
rate.
Another idea: pharming putting genes for useful
proteins like insulin into plants, and letting the plant
synthesize them in large amounts. Can also be
done with animals: proteins secreted into milk.

Nuclear Cloning

Why not just take the nucleus


from any cell and put it into an
egg, producing a new person
genetically identical to the
original? This is the idea
behind nuclear cloning, and it
does work on occasion.
Dolly the Sheep was the first
example: a nucleus from her
parents mammary gland was
extracted and put into an egg
whose own nucleus was
removed. The egg was them
implanted into the uterus of
another sheep, and Dolly was
born. She is genetically
identical to the donor sheep.

More Nuclear Cloning

Although a clones nuclear DNA is identical to the donor parent, the


parent and offspring are not likely to be exactly identical. Similarly,
identical twins are not exactly identical. Events occur after the
identical zygotes form: random environmental influences, patterns of
development, etc.
It is very difficult to get a nucleus of a cell to regress to the
totipotent state of an embryonic cell. A totipotent cell can turn into
any cell type. After a few cell divisions of the embryo, the cells are
restricted: they can no longer become any type. Many genes are
permanently inactivated by modifying the DNA molecule. Removing
these modifications is, at the moment, more of an art than a
science.
The result: the failure rate in cloning is very high: less than 1%
success rate. And, of the clones that are born alive, many suffer
defects that only become apparent later in life. Dolly, for instance,
died at a young age.