BIOB11H3 Lecture 2: Lecture 2

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4 Jul 2018

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Lecture 2

The Structure of The Genome

Genome: The entire genetic makeup of an organism i.e. all the genetic information that is present in

an organism

Eukaryotic cells: DNA present in the nucleus, mitochondria, chloroplasts, plastids and maybe some

extra-chromosomal DNA (outside of nucleus)

Prokaryotic cells: DNA present in the chromosome

Viruses: Double or single stranded DNA or RNA

Humans: Essentially all the genetic information is in

a single haploid set of chromosomes – 22

autosomes (chromosomes that are not sex

chromosomes) and both the X and Y sex

chromosomes.

Human genome is much more complex than that of

viruses and bacteria.

DNA Denaturation and Renaturation

The ability of the two DNA strands to unwind and separate into two individual strands is called

denaturation

Based on the fact that the double stranded (ds) DNA molecule (helix) is held together by hydrogen

bonds (weak and non-covalent) between the bases.

Complementary single stranded DNA molecules can re-associate – Renaturation or Reannealing

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DNA Denaturation

Experimentally when DNA is dissolved in a weak saline solution and slowly warmed to a certain

temperature DNA strand separation begins. Within a few degrees (°C) of initiation of strand separation,

the two chains are completely separated from each other (driven apart by electrostatic repulsion of

the negative charges on the sugar-phosphate backbone)

Thermal denaturation of DNA = DNA Melting

Different DNA sequences denature at different temperatures

The progress of thermal DNA denaturation (melting) is monitored by following the increase in

absorbance of the dissolved DNA.

Thermal Denaturation of DNA

The nitrogenous bases of a nucleic acid absorb UV radiation (maximum at 260 nm)

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260 nm wavelength can be used to determine DNA

concentration

Single stranded (ss) DNA absorbs almost twice as much

UV light as compared to double stranded (ds) DNA.

Melting Curve: UV absorbance on Y-axis, temperature on

X-axis

Tm: the temperature at which the shift in absorbance is

half completed

Originally thought to be solely due to the number of

hydrogen bonds that hold G/C base pairs together versus

A/T pairs

It also has to do with the strength of base-stacking

interactions (figure G)

G/C pairs have stronger stacking interactions than A/T pairs

The higher the GC content of DNA, the higher the Tm due to the extra hydrogen bond between the

G-C pairs

DNA Renaturation

Complementary single strands of DNA can

re-associate (reanneal) into a stable

double stranded DNA molecule

Occurs when denatured DNA is allowed to

cool slowly – complementary strands

‘find’ each other and base pairing begins

After renaturation, DNA becomes a

double-stranded helical molecule

Example: Comparing the rate of

renaturation of different sized genomes

(C0t plots)

Example of DNA in genome that is

predominately single copy (no significant amount of repeated DNA, one gene after another)

C0t Plots (Kinetics of Renaturation)

C0t values:

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Document Summary

Genome: the entire genetic makeup of an organism i. e. all the genetic information that is present in an organism. Eukaryotic cells: dna present in the nucleus, mitochondria, chloroplasts, plastids and maybe some extra-chromosomal dna (outside of nucleus) Viruses: double or single stranded dna or rna. Humans: essentially all the genetic information is in a single haploid set of chromosomes 22 autosomes (chromosomes that are not sex chromosomes) and both the x and y sex chromosomes. Human genome is much more complex than that of viruses and bacteria. The ability of the two dna strands to unwind and separate into two individual strands is called denaturation. Based on the fact that the double stranded (ds) dna molecule (helix) is held together by hydrogen bonds (weak and non-covalent) between the bases. Complementary single stranded dna molecules can re-associate renaturation or reannealing. Experimentally when dna is dissolved in a weak saline solution and slowly warmed to a certain temperature dna strand separation begins.

Related Questions

In addition to identifying the genetic material, the experiments of Avery, MacLeod, and McCarty with different strains of Streptococcus pneumoniae demonstrated that

	DNA is a double helix.
	DNA may be taken up by bacterial cells and be active.
	DNA is present in bacteriophage T2.
	eukaryotic cells may be genetically transformed.
	DNA binds to the cell wall of the bacterium.

In order to show that DNA in cell extracts is responsible for genetic transformation in Streptococcus pneumoniae, important corroborating evidence should indicate that _______ also destroy transforming activity.

	temperatures high enough to kill cells
	enzymes that hydrolyze proteins
	enzymes that hydrolyze DNA
	enzymes that hydrolyze polysaccharides
	enzymes that hydrolyze RNA

Based on what you have learned about the experiments conducted by Griffith and Avery and colleagues with bacteria, which of the following would result in transformation of living R cells?

	Pure RNA from S cells
	Cell-membrane extracts from S cells
	A mixture of pure RNA and protein from S cells
	Cell-free extracts from S cells
	Pure protein from S cells

A-T base pairs in a DNA double helix

	form three hydrogen bonds with each other.
	are more heat-stable than G-C base pairs.
	are of a substantially different length than G-C base pairs.
	are not accessible to DNA binding proteins.
	are chemically distinct from G-C base pairs.

If 23 percent of the bases in a sample of double-stranded DNA are adenine, what percentage of the bases are uracil?

	27
	50
	0
	73
	23

The uniform diameter of the DNA structure provides evidence for

	antiparallel DNA strands.
	a right-handed helix.
	a double-stranded helix.
	3â² end phosphorylation.
	purineâpyrimidine pairing.

If a sequence of one strand of DNA is 5â²-TGACTATC-3â², what is the complementary strand?

	5â²-ACTGATAG-3â²
	3â²-TGACTATC-5â²
	3â²-ACTGATAG-5â²
	3â²-ACTGATAG-3â²
	5â²-TGACTATC-3â²

What structural aspect of the DNA facilitates dissociation of the two DNA strands for replication?

	3â² end phosphorylation
	Purineâpyrimidine pairing
	Antiparallel DNA strands
	Covalent bonds between sugars and phosphate groups
	Hydrogen bonding between paired bases

If the MeselsonâStahl density gradient experiment had resulted in two bands of DNA molecules after only one round of replication, one containing only ¹⁵N and the second only ¹⁴N, this result would have indicated that replication was

	unidirectional.
	dispersive.
	conservative.
	bidirectional.
	semiconservative.

10.

The nucleoside analogue acyclovir, which is used to treat herpes simplex virus (HSV) infections, lacks a 3â² hydroxyl group (âOH). Predict what will happen if the host cell DNA polymerase incorporates a molecule of acyclovir into an elongating strand of HSV DNA.

	The replication complex will no longer bind to origins of replication.
	The DNA polymerase will only incorporate acyclovir molecules.
	Acyclovir will bind single-strand binding proteins.
	DNA polymerase will not be able to link a successive nucleotide.
	Acyclovir will block the activity of the DNA helicase.

11.

Which of the following does not demonstrate the stability of the DNA double helix?

	Single-strand binding proteins are required to keep the strands apart.
	Paired bases form hydrogen bonds.
	High temperatures are required to denature it.
	DNA synthesis requires energy.
	DNA helicases require ATP.

12.

What effect would a primase inhibitor have on DNA replication?

	DNA polymerase would not be able to add bases to the DNA.
	Primase would be blocked from joining the DNA replication complex.
	Primase would not be able to provide primers for DNA polymerases.
	DNA polymerase would not be able to use DNA as a template.
	There would be no effect on DNA replication.

13.

To replicate their DNA in a timely manner, most eukaryotic chromosomes

	normally have uncoiled DNA.
	have multiple origins of replication.
	contain linear molecules of single-stranded DNA.
	have two replication forks that move in the same direction.
	are composed entirely of DNA.

14.

Which statement about DNA replication is false?

	Okazaki fragments are synthesized as parts of the leading strand.
	Error rates for DNA replication are reduced by proofreading of the DNA polymerase.
	The sliding clamp increases the rate of DNA synthesis.
	Replication forks represent areas of active DNA synthesis on the chromosomes.
	Ligases and polymerases function in the vicinity of replication forks.

15.

In many eukaryotes, there are repetitive sequences called telomeres at the ends of chromosomes. After successive rounds of DNA replication, the _______ strand becomes shorter. In some cells, an enzyme called _______ repairs the shortened strand.

	leading; telomerase
	lagging; telomerase
	lagging; DNA polymerase I
	leading; DNA polymerase I
	leading; replicase

16.

A researcher studies normal human fibroblast cells. They can be maintained in culture but die off after about 30 cell generations. Unexpectedly, a colony of cells continues to survive and divide past 30 generations. Which scenario is most likely true for these cells?

	DNA polymerase is constantly active.
	Primase is able to extend the telomeres.
	The mismatch repair system is extra efficient.
	The telomerase gene is being expressed.
	The cells do not need the ends of their chromosomes.

17.

If DNA polymerase III introduces an incorrect nucleotide, what is the first corrective action made by the DNA repair system?

	The replication complex excises the incorrect nucleotide.
	The excision repair proteins remove the incorrect nucleotide.
	The mismatch repair system scans for base-pairing mismatches.
	There is no correction, because nucleotide errors by DNA polymerases are not repaired.
	DNA ligase connects the correct nucleotide.

18.

Choose the correct order of the following four events in the excision repair of DNA:
(1) Base-paired DNA is made complementary to the template.
(2) Damaged bases are recognized.
(3) DNA ligase seals the new strand to existing DNA.
(4) Part of a single strand is excised.

	1, 2, 3, 4
	2, 1, 3, 4
	3, 4, 2, 1
	2, 4, 1, 3
	4, 2, 3, 1

19.

Six complete cycles of PCR should result in a _______-fold increase in the amount of DNA.

	64
	32
	12
	128
	6

20.

When double-stranded DNA is heated to temperatures above 90Â°C, it denatures. Denaturation is a process that

	breaks covalent bonds.
	hydrolyzes DNA.
	separates double-stranded DNA into two single strands.
	causes new hydrogen bonds to form.
	irreversibly destroys the double helical structure.

QUESTION 1: The rabies virus primarily affects the nervous system. The specificity that the rabies virus has for neuronal host cells is primarily dictated by __________________________.

A. the helical shape of its viral capsid

B. the type of nucleic acid used for its viral genome (single-stranded, antisense RNA)

C. the spikes that protrude from its viral envelope

D. the segmented nature of its viral genome

QUESTION 2: Which of the following genome types has been observed in viruses? To be marked correct, you'll need to select all true statements, as there may be more than one correct answer.

A. Single-stranded RNA

B. Single-stranded DNA

C. Double-stranded RNA

D. Protein-based

QUESTION 3: Choose the correct statement about viral evolution.

A. RNA and DNA viral genomes evolve at equal rates.

B. Cellular genomes mutate at a faster rate than viral genomes due to their large size and increased chance of replicative mistakes.

C. DNA viruses mutate faster than RNA viruses because thymine is more susceptible to mutation than uracil.

D. RNA viruses mutate faster than DNA viruses due to a lack of proofreading replicative enzymes.

QUESTION 4: Your elderly patient is affected by shingles. After careful observation, you note that the virus responsible for the infection has an icosahedral capsid, is enveloped, and has double-stranded linear DNA as its genetic material. Based on this information, in which of the following viral families would you group this viral pathogen?

QUESTION 5: Which of the following is a key difference between lytic and lysogenic bacteriophage replication cycles?

A. Lysogenic bacteriophages release via host cell lysis, whereas lytic bacteriophages bud off from the host cell membrane.

B. The lytic replication cycle ends with host cell lysis and release of newly formed bacteriophage particles, whereas lysogenic replication leads to prophage formation.

C. Lytic replication requires an uncoating step, whereas in lysogenic replication, the bacteriophage's nucleic acid is directly injected into the host cell.

D. Whereas lytic replication enables the infection of several hosts, lysogenic replication only allows for infection of a single host cell.

QUESTION 6: Which of the following is an unsuitable culture method for an animal virus?

A. Direct inoculation of virus onto blood agar

B. Live animal inoculation

C. Tissue culture method

D. Injection of virus into embryonated eggs

What step in DNA replication precedes the pairing of complementary bases?

	A.	polymerization of DNA.
	B.	formation of the histone core.
	C.	mitotic division.
	D.	separation of the two strands.
	E.	joining of the two strands.

In eukaryotes, transcription occurs in the

	A.	ribosome.
	B.	mitochondria.
	C.	cell membrane.
	D.	cytoplasm.
	E.	nucleus.

Normal human body cells contain how many chromosomes?

	A.	46
	B.	22
	C.	23
	D.	44
	E.	42

Hershey and Chase performed an experiment in which they used radioactive isotopes to phosphorus and sulfur to label different components of bacterial viruses. Their results clearly showed that

	A.	the viruses did not contain protein.
	B.	genetic material normally contains radioactive isotopes.
	C.	the viruses did not contain DNA.
	D.	protein was entering into the infected bacteria.
	E.	DNA was entering into the infected bacteria.

A package of histones with DNA wrapped around them forms spherical structures called

	A.	nucleosomes.
	B.	lysosomes.
	C.	ribosomes.
	D.	nucleoli.
	E.	chromatin.

DNA replication

	A.	does not require proteins.
	B.	is constantly happening in a cell.
	C.	occurs in G1 of interphase.
	D.	takes place in the nucleus of the cell.
	E.	occurs in the cytoplasm of the cell.

The two subunits of the ribosome join during

	A.	promotion of transcription.
	B.	initiation of translation.
	C.	termination of transcription.
	D.	termination of translation.
	E.	elongation.

DNA replication of a single DNA molecule is referred to as semiconservative because

	A.	each of the two DNA molecules will consist of one parental strand and one newly synthesized strand.
	B.	all the DNA strands in the two DNA molecules will have both parental and newly synthesized DNA.
	C.	it results in two nonidentical DNA molecules.
	D.	of the two DNA molecules, one is made up totally of parental DNA, while the other is entirely newly synthesized DNA.
	E.	one DNA strand is replicated continuously, while the other must be replicated discontinuously.

The bases are bonded to what part of the backbone of the DNA molecule?

	A.	the 4â carbon atom of the sugar molecule.
	B.	the 3â carbon atom of the sugar molecule.
	C.	the 5â carbon atom of the sugar molecule.
	D.	the 1â carbon atom of the sugar molecule.
	E.	the 2â carbon atom of the sugar molecule.

BIOB11H3 Lecture 2: Lecture 2

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