GENERAL BIOLOGY 2
Chapter 9 the mice became fatally ill, and their blood
DNA Structure and Function contained live S cells.
9.1 A Hero Dog’s Golden Clones Avery and McCarty Find the Transforming Principle
• James Symington and his search dog Trakr located • 1940: Avery and McCarty separated deadly S cells
the last living survivor of the 9/11 attack on the (from Griffith’s experiments) into lipid, protein, and
World Trade Center nucleic acid components
• Trakr later died of a degenerative neurological • When lipids, proteins, and RNA were destroyed, the
disease, probably due to toxic smoke exposure at remaining substance, DNA, still transformed R cells
Ground Zero to S cells
• Trakr’s DNA lives on in his clones – genetic copies • Conclusion: DNA is the “transforming principle”
produced by inserting his DNA into donor eggs
Confirmation of DNA’s Function
The Cloning Controversy • 1950s: Hershey and Chase experimented with
• Few cloned mammal embryos result in a live birth – bacteriophages (viruses that infect bacteria)
many of the clones that survive have serious health - Protein parts of viruses, labeled with 35S, stayed
problems outside the bacteria
- The problem: DNA in adult cells is controlled - DNA of viruses, labeled with 32P, entered the
differently than the DNA in embryonic cells bacteria
• Perfecting methods for cloning animals bring us • Conclusion: DNA, not protein, is the material that
closer to the possibility of cloning humans, both stores hereditary information
technically and ethically
*CLONIING: ASEXUAL REPRODUCTION Bacteriophages
Figure 9.3 The Hershey–Chase experiments. Alfred
9.2 The Discovery of DNA’s Function Hershey and Martha Chase carried out experiments to
• 1869: Johannes Miescher found DNA determine the composition of the hereditary material that
(deoxyribonucleic acid) in nuclei, though its function bacteriophage inject into bacteria. The experiments were
was unknown based on the knowledge that proteins contain more sulfur (S)
• Early 1900s: Griffith transferred hereditary material than phosphorus (P), and DNA contains more phosphorus
from dead cells to live cells than sulfur.
- Mice, injected with live R cells, lived
- Mice, injected with live S cells, died
- Mice, injected with killed S cells, lived
- Mice, injected with killed S cells and live R cells,
died; live S cells were found in their blood
*PURPOSE OF CLONING:
- Therapy
- Increase number of Endangered Species
Griffith’s Experiment
A. Left, a model of a bacteriophage. Right, micrograph
of three viruses injecting DNA into an E. coli cell.
Figure 9.2 Fred Griffith’s experiments with two strains (R and
S) of Streptococcus pneumoniae bacteria.
A. Griffith’s first experiment showed that R cells were B. In one experiment, bacteriophage was labeled with a
harmless. When injected into mice, the bacteria radioisotope of sulfur (35S), a process that makes
multiplied, but the mice remained healthy. their protein components radioactive. The labeled
B. The second experiment showed that an injection of S viruses were mixed with bacteria long enough for
cells caused mice to develop fatal pneumonia. Their infection to occur, and then the mixture was whirled
blood contained live S cells. in a kitchen blender. Blending dislodged viral parts
C. For a third experiment, Griffith killed S cells with that remained on the outside of the bacteria.
heat before injecting them into mice. The mice Afterwards, most of the radioactive sulfur was
remained healthy, indicating that the heat-killed S detected outside the bacterial cells. The viruses had
cells were harmless. not injected protein into the bacteria.
D. In his fourth experiment, Griffith injected a mixture
of heat-killed S cells and live R cells. To his surprise,
• This was the final piece of information James
Watson and Francis Crick needed to build their
model of DNA
C. In another experiment, bacteriophage was labeled Watson and Crick’s DNA Model
with a radioisotope of phosphorus (32P), which • A DNA molecule consists of two nucleotide chains
makes their DNA radioactive. The labeled viruses (strands), running in opposite directions and coiled
were allowed to infect bacteria. After the external into a double helix
viral parts were dislodged from the bacteria, the • Base pairs form on the inside of the helix, held
radioactive phosphorus was detected mainly inside together by hydrogen bonds (A-T and G-C)
the bacterial cells. The viruses had injected DNA into
the cells—evidence that DNA is the genetic material Rosalind Franklin, X-Rays, and Cancer
of this virus. • In science, as in other professions, public recognition
does not always include everyone who contributed
9.3 The Discovery of DNA’s Structure to a discovery
• Nucleotide • Rosalind Franklin was first to discover the
- A nucleic acid monomer consisting of a five- molecular structure of DNA, but did not
carbon sugar (deoxyribose), three phosphate groups, share in the Nobel prize which was given to
and one of four nitrogen-containing bases Watson, Crick, and Wilkins
• DNA consists of four nucleotide building blocks • Franklin died of cancer at age 37, probably
- Two pyrimidines: thymine and cytosine caused by extensive exposure to x-rays
- Two purines: adenine and guanine during her work
Four Kinds of Nucleotides in DNA 9.4 Eukaryotic Chromosomes
• The DNA in a eukaryotic cell nucleus is organized as
one or more chromosomes that differ in length and
shape
• Chromosome
• A structure that consists of DNA and
associated proteins
• Carries part or all of a cell’s genetic
information
Chromosome Structure – Illustrated
Figue 9.4 The four nucleotides in DNA. Each has three
phosphate groups, a deoxyribose sugar (orange), and a
nitrogen-containing base (blue) after which it is named.
Adenine and guanine bases are purines; thymine and
cytosine, pyrimidines. Biochemist Phoebus Levene identified
the structure of these bases and how they are connected in
nucleotides in the early 1900s. Levene worked with DNA for
almost 40 years.
Chargaff’s Rules
• The amounts of thymine and adenine in DNA are the Figure 9.7 Chromosome structure
same, and the amounts of cytosine and guanine are 1. Two strands of DNA twist into a double helix.
the same 2. At regular intervals, the DNA (blue) wraps around a
- A=T core of histone proteins (purple).
- G=C 3. The DNA and proteins associated with it twist tightly
• The proportion of adenine and guanine differs into a fiber.
among species 4. The fiber coils and then coils again to form a hollow
cylinder.
Franklin, Watson, and Crick 5. At its most condensed, a duplicated chromosome
• Rosalind Franklin’s research in x-ray crystallography has an X shape.
revealed the dimensions and shape of the DNA 6. The DNA in the nucleus of a eukaryotic cell is
molecule: an alpha helix typically divided into a number of chromosomes.
*She was the one who found DNA
Chromosome Number • DNA polymerase joins free nucleotides into a new
• The total number of chromosomes in a eukaryotic strand of DNA
cell (chromosome number) is characteristic of the • DNA ligase joins DNA segments on the discontinuous
species strand
• Human body cells have:
- Forty-six chromosomes Primers for DNA Polymerase
- Two of each type of chromosome – so their • Several types of DNA polymerases exist
chromosome number is diploid (2n) • Each requires a primer to initiate DNA
• A karyotype shows how many chromosomes are in synthesis
an individual cell, and reveals major structural • A primer is a short, single strand of DNA or
abnormalities RNA that is complementary to a targeted
DNA sequence
Semiconservative DNA Replication
• Each strand of a DNA double helix is a template for
synthesis of a complementary strand of DNA
• One template builds DNA continuously; the other
builds DNA discontinuously, in segments
• Each new DNA molecule consist of one old strand
and one new strand (semiconservative replication)
Figure 9.9 Karyotypes
A Karyotype of a female human, with identical sex Figure 9.10 DNA replication, in which
chromosomes (XX) a double-stranded molecule of DNA
is copied in its entirety. green arrows
Autosomes and Sex Chromosomes show the direction of synthesis for
• In a diploid organism, one chromosome in a pair is each strand. The Y-shaped structure
inherited from the mother and one from the father of a DNA molecule undergoing
- All except one pair of chromosomes are replication is called a replication fork.
autosomes – chromosomes with the same
length, shape, and centromere location 1. As replication begins, many
- Pairs of sex chromosomes differ between initiator proteins attach to
females and males – human females have two X the DNA at certain sites in
chromosomes (XX); human males have one X the chromosome.
and one Y chromosome (XY) Eukaryotic chromosomes have many
of these origins of replication; DNA
9.5 DNA Replication replication proceeds more or less
• DNA replication is the energy-intensive process by simultaneously at all of them.
which a cell copies its DNA 2. Enzymes recruited by the initiator proteins begin to
• A cell copies its DNA before it reproduces unwind the two strands of DNA from one another.
• Each of the two DNA strands in the double helix is 3. Primers base-paired with the exposed single DNA
replicated strands serve as initiation sites for DNA synthesis.
• DNA replication requires many enzymes, including 4. Starting at primers, DNA polymerases (green boxes)
DNA polymerase, and other molecules assemble new strands of DNA from nucleotides,
using the parent strands as templates.
Replication of the DNA Sequence 5. DNA ligase seals any gaps that remain between
• A cell’s genetic information consists of the order of bases of the “new” DNA, so a continuous strand
nucleotide bases (the DNA sequence) of its forms.
chromosomes 6. Each parental DNA strand (blue) serves as a template
• Descendant cells must get an exact copy of that for assembly of a new strand of DNA (magenta).
information Both strands of the double helix serve as templates,
• Each chromosome is copied entirely – the two so two double- stranded DNA molecules result. One
chromosomes that result are duplicates of the strand of each is parental (old), and the other is new,
parent molecule so DNA replication is said to be semiconservative.
Enzymes of DNA Replication Discontinuous Replication
• DNA helicase breaks hydrogen bonds between DNA • DNA polymerases attach a free nucleotide only to
strands the 3′ end of a DNA strand
• Topoisomerase untwists the double helix
• Only one of the two new strands of DNA can be - Egg cytoplasm reprograms differentiated (adult)
synthesized continuously during DNA replication DNA to act like undifferentiated (egg) DNA
• Synthesis of the other strand occurs in segments, in - The hybrid cell develops into an embryo that is
the direction opposite that of unwinding genetically identical to the donor individual
• DNA ligase joins segments into a continuous strand
of DNA
Figure 9.13 Somatic cell nuclear transfer, using cattle cells.
This series of micrographs was taken by scientists at Cyagra, a
company that specializes in cloning livestock.
Therapeutic Cloning
• Therapeutic cloning uses SCNT to produce human
embryos for research purposes
• Researchers harvest undifferentiated (stem) cells
from the cloned human embryos
• Such research may ultimately lead to treatments for
people who suffer from fatal diseases
9.8 Cloning DNA
• Restriction enzymes
- Bacterial enzymes that cut DNA wherever a
specific nucleotide sequence occurs
Figure 9.11 Discontinuous synthesis of DNA. This close-up of a • Single-stranded DNA tails produced by the same
replication fork shows that only one new DNA strand is restriction enzyme base-pair together
assembled continuously. - DNA ligase bonds “sticky ends” together
• Recombinant DNA
9.6 Mutations: Cause and Effect - Composed of DNA from two or more organisms
• DNA polymerases proofread DNA sequences during
DNA replication and repair damaged DNA Making Recombinant DNA
• When proofreading and repair mechanisms fail, an Figure 9.15 Making recombinant DNA
error becomes a mutation – a permanent change in
the DNA sequence
9.7 Cloning Adult Animals
• Clones
- Exact copies of a molecule, cell, or individual
- Occur in nature by asexual reproduction or
embryo splitting (identical twins) 1. A restriction enzyme recognizes a specific base
• As cells develop, they become differentiated sequence (orange boxes) in DNA from any source.
- Different in form and function
- Usually a one-way process in animal cells
• Reproductive cloning technologies produce an exact
copy (clone) of an individual
Cloning in the Laboratory
• Somatic cell nuclear transfer (SCNT)
- Nuclear DNA of an adult is transferred to an
enucleated egg 2. The enzyme cuts DNA from two sources into
fragments. This enzyme leaves sticky ends
The Human Genome Project
• Automated DNA sequencing and PCR allowed human
genome projects to sequence the 3 billion bases in
the human genome
- 28,976 genes have been identified, but not all of
their products or functions are known
3. When the DNA fragments from the two sources are
mixed together, matching sticky ends base-pair with A Human DNA Sequence
each other.
4. DNA ligase joins the base-paired DNA fragments.
Molecules of recombinant DNA are the result.
DNA Cloning Figure 9.21 Human DNA sequence data. The order of colors in
• Making recombinant DNA is the first step in DNA each vertical lane reveals one part of the DNA sequence.
cloning, a set of laboratory methods that uses living
cells to mass-produce specific DNA fragments Genomic DNA Alignment
- DNA cut into fragments by restriction enzymes
is inserted into cloning vectors (plasmids) cut
with the same enzyme
- Cloning vectors with foreign DNA are placed in Figure 9.23 Genomic DNA alignment. This is a region of the
host cells, which divide and produce many gene for a DNA polymerase. Nucleotides that differ from
clones, each with a copy of the foreign DNA those in the human sequence are highlighted. The chance
that any two of these sequences would randomly match is
cDNA Cloning about 1 in 1046.
• Complementary DNA (cDNA)
- DNA made from an mRNA template
• Reverse transcriptase transcribes mRNA to DNA,
forming a hybrid molecule
- DNA polymerase builds a double-stranded DNA
molecule that can be cloned
9.9 Isolating Genes
• Genome
- The entire set of genetic material of an organism
• DNA libraries are sets of cells containing various
cloned DNA fragments
- Genomic libraries (all DNA in a genome)
- cDNA libraries (all active genes in a cell)
