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28 Nov 2018

1.) Choose the best response for each.

a.) The Ames test is commonly used to test whether a particular chemical is mutagenic. When you conduct an Ames test, you need two groups of plates. One group of plates contains bacteria that are exposed to the chemical of interest, while the other plates contain bacteria that are not exposed to the chemical. Why is the latter group needed if you are just interested in the rate of mutation caused by the chemical?

The group without the chemical can be exposed to mammalian liver extract to see the effects of liver enzymes on the bacteria.

Having two groups allows researchers to compare the effects of different environments on the mutation rate

It is important to compare the rate of mutations with the chemical to the normal background mutation rate. Mutations occur spontaneously even without the presence of a mutagen.

The second group is used as a control to make sure that the replica plating technique is done correctly.

b.) Frameshift mutations can be very severe. What is one reason why they are often so serious?

When a frameshift mutation occurs, it causes transition mutations to occur. These are particularly severe.

When a frameshift mutation occurs, it causes changes in many bases downstream and can affect many of the amino acids in the protein.

Frameshift mutations always occur in the regulatory region, so they cause a gene to be turned off and then the protein is not produced at all.

When a frameshift mutation occurs, it causes changes in many bases upstream and can affect many of the amino acids in the protein.

c.) An organism that cannot synthesize methionine is called __________.

a mutant

a prototroph

an auxotroph

an autotroph

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Bunny GreenfelderLv2

29 Nov 2018

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1. At Thanksgiving dinner, your great aunt refuses to eat with plastic silverware and proudly exclaims that she has discarded all plastics because âthey cause cancer.â You wonder if chemicals you commonly use are mutagenic. You recall your time in BY 331, Genetics, and realize that you can use the Ames test to test for reversion of a point mutation in the HisG gene in the bacteria S. typhimurium. You added the chemical you wish to test into the growth medium for the bacteria. You carefully plated an equal number of cells on each mutagenesis plate. You also calculated percent survival for chemical treated cells relative to untreated cells. Remember that the plate media for survival is not selective. A summary of the results is shown below.

Chemical	Percent Survival	Number of Revertants (# of His⁺ colonies/selective plate)
No chemical added	100	28
Chemical A	50	1400
Chemical B	70	20
Chemical C	100	7

A. What medium must be used in the selective plate as part of the Ames test? Explain how a mutation gives rise to a revertant in this experiment.

[Type your answer here, use as much space as necessary to completely answer the question]

B. You are initially surprised to see revertants in the absence of any chemical that you are testing, but you realize that this is normal. Explain a specific way in which a revertant can arise in the absence of an added mutagen.

[Type your answer here, use as much space as necessary to completely answer the question]

C. Which chemicals would you identify as containing a mutagen? Explain your reasoning.

[Type your answer here, use as much space as necessary to completely answer the question]

D. Which chemicals would you identify as possibly antimutagenic? Explain your reasoning.

[Type your answer here, use as much space as necessary to completely answer the question]

QUESTION 1

A mutation caused by exposure to gamma rays is called a spontaneous mutation

	a.	True
	b.	False
	c.	Sometimes, depending on the type of base change
	d.	Gamma rays do not cause mutations

QUESTION 2

A recessive mutation affecting an essential biochemical pathway can be detected________

	a.	when the organism is heterozygous for that mutation.
	b.	when the organism is homozygous for that mutation.
	c.	in either homozygous or heterozygous conditions.
	d.	only by sequencing (recessive mutations never show a phenotype).

QUESTION 3

During DNA replication, the newly synthesized chain grows by adding

	a.	nitrogenous bases
	b.	the sugar-phosphate backbone
	c.	nucleosides
	d.	dNTPs

QUESTION 4

During replication, the DNA strand 5â- AAGTCTAGCCTAG -3â will serve as a template for the polymerization of:

	a.	5â- CTAGGCTAGACTT -3â
	b.	5â - TTCAGATCGGATC -3â
	c.	5â - GATCCGATCTGAA -3â
	d.	3â - AAGTCTAGCCTAG -5â

QUESTIOn 5

Given the following DNA sequences, select which statement(s) is(are) correct.

Sequence 1:

5â-TGGACGCTAA-3â

3â-ACCTGCGATT-5â

Sequence 2:

5â-AATCGCAGGT-3â

3â-TTAGCGTCCA-5â

Sequence 3:

5â-ACCTGCGATT-3â

3â-TGGACGCTAA-5â

	a.	Sequences 1 and 2 are the same
	b.	Sequences 1 and 3 are the same
	c.	Sequences 2 and 3 are the same
	d.	Sequences 1, 2 and 3 are all different

QUESTION 6

Repetitive sequences in the genome are hotspots for:

	a.	Deamination
	b.	Depurination
	c.	Thymine dimer formation
	d.	Replication errors

QUESTION 7

Select which statement(s) about the Ames test is(are) correct

	a.	it has been designed to understand mutation repair systems in Salmonella typhimurium
	b.	it uses mammalian liver extract
	c.	it allows to study whether chemical compounds or their enzymatic breakdown products are mutagenic
	d.	it is based on whether a chemical compound causes reversion mutations from his+ to his-
	e.	two of the above are correct
	f.	three of the above are correct

QUESTION 8

Select which statement(s) is(are) correct.

	a.	All DNA strands have a direction, and it is specified by the carbons in the sugar backbone.
	b.	All DNA strands have a direction, and it is specified by hydrogen bonds between nucleotides.
	c.	In order to form proper base pairs in a double stranded DNA molecule, the two strands must run in opposite directions.
	d.	A and C are correct.
	e.	B and C are correct.

QUESTION 9

Studies of gene mutation frequencies have shown that:

	a.	mutations are rare, and genomes are generally stable.
	b.	mutation frequencies differ among organisms and also between genes, suggesting certain genes are more susceptible to mutation.
	c.	mutation frequencies are consistent between organisms, and each region of DNA is equally susceptible to random mutations.
	d.	Both A and B are correct.
	e.	Both A and C are correct.

QUESTION 10

The compound 5-bromodeoxyuridine (BrdU) is a derivative of uracil, and if BrdU becomes incorporated during DNA replication, it pairs with adenine. This compound is best classified as which type of mutagen?

	a.	base analog
	b.	base inducer
	c.	intercalating agent
	d.	oxidative agent
	e.	alkylating agent

QUESTION 11

The rate of mutation of the fruit fly is higher than the rate of mutation of Algae

True

False

QUESTION 12

Thymine dimers are most commonly caused by which of the following?

	a.	X-rays
	b.	Alkylating agents
	c.	U.V. irradiation
	d.	DNA intercalating agents

QUESTION 13

What chemical group is found in the 3â end of a DNA strand?

	a.	an alcohol
	b.	a hydroxyl
	c.	a methyl
	d.	a phosphate

QUESTION 14

What chemical group is found in the 5â end of a DNA strand?

	a.	an alcohol
	b.	a hydroxyl
	c.	a methyl
	d.	a phosphate

QUESTION 15

Which is the correct order of molecules binding to DNA during DNA replication?

	a.	Helicase, SSB, primase, DNA pol III, DNA pol I, ligase
	b.	SSB, DNA pol I, ligase, helicase, DNA pol I, primase
	c.	primase, helicase, DNA pol III, ligase, DNA pol I, SSB
	d.	SSB, helicase, primase, DNA pol III, DNA pol I, ligase

QUESTION 16

Which statement(s) is(are) correct about strand slippage?

	a.	It can cause disorders such as Huntingon disease.
	b.	It is a process that causes mutations altering the number of DNA repeats.
	c.	It is a process that incorporates nucleotide base analogs and trinucleotide repeats.
	d.	a and b are correct.
	e.	b and c are correct.
	f.	a, b and c are correct.

help needed asap.

chocolatesea-lion434

For Discussion (2-3 paragraphs) â¢ What was the concentration of your pre-diluted NPs? â¢ Determine the effect of NPs on brine shrimp lethality. â¢ Determine the effect of NPs on cell viability. â¢ What is (are) the control(s) in this experiment? What is its purpose? â¢ Based on your data in this lab, what is a safe concentration for brine shrimp Threshold Level of Toxicity (TTL)? â¢ Often indicator species are used to study the overall health of an ecosystem. If you were to study an ecosystem containing brine shrimp, would you use it as an indicator species? Why or Why not? Explain your reasoning. â¢ What possible sources of error were present in this experiment?

Introduction

LD50 is a commonly used term to describe acute toxicity: LD stands for Lethal Dose and 50 is used because the dose could kill 50% of the animals exposed to the chemical. For LD50 tests, the animals are given specific amounts of the chemical, orally, by injection or dermally. The value is then reported along with the method of administration and the type of animal used in the test. LD50 is typically reported in milligrams or grams per kilogram of body weight of the animal. The lower the LD50 is, the higher the toxicity of the chemical would be. This information is always provided in Safety Data Sheet (SDS) of chemicals in the âtoxicity dataâ category. A similar measure, the LC50 is usually used to describe the ecological hazards of chemicals. LC stands for lethal concentration that could kill 50% of the animals after 24-96 hours. This value is always provided in Safety Data Sheet (SDS) of chemicals in the âEcological Dataâ category. In this lab, you will be measuring LC50 or LD50 of iron oxide nanoparticles using brine shrimp. Iron oxide nanoparticles are magnetic. Application of magnetic nanoparticles ranges from magnetic storage media to medical industry where they can be used for targeted drug delivery or as contrast agents in MRI. Thus, measuring environmental and biological toxicity of these nanoparticles is crucial. To test toxicity of any such reagents on small organisms, Daphnia and brine shrimps (also known as Artemia) can be used. Artimia are found in salt lakes around the world. They are important live food for aquarium fish. In this lab, you will use brine shrimp to measure toxicity of the nanoparticles. Trypan blue is a stain used to assess cell viability. It is able to diffuse through cell membranes of dead cells but is unable to permeate live cells. This stain can be used to assess cell survival 1 Fall 2017 following exposure to chemicals, nanoparticles and experimental drugs. The ratio of live: dead cells can be used as a preliminary assessment of cytotoxicity and translated to tissue viability.

Methods Part A: Serial Dilution of NP Solution 1) Label 5 test tubes as 0, 1:1, 1:10, 1:100, 1:1000 2) Add 2mL of seawater to tube 0 3) Add 2mL of specific group stock solution to tube 1:1 and add 2mL of seawater to it. 4) Transfer 1mL of stock solution to tube 1:10 and add 9mL of seawater to this tube 5) Transfer 1mL of 1:10 solution to the tube labeled as 1:100 and add 9mL of seawater to â¨this tube 6) Transfer 1mL of 1:100 solution to the test tube labeled as 1:1000 and add 9mL of â¨seawater to this tube Part B: Preparation of test environments 1) Label 5 wells of 6-well plate as control (0), 1:1, 1:10, 1:100 and 1:1000 2) Label 3 petri dishes as: control (0), 1:1 and 1:1000 â¨NOTE: Be sure to include your initials on all plates 3) Transfer 2mL of each solution from the corresponding test tube to the respective wells on your six-well plate and each petri dish Part C: Preparing specimens for the Trypan blue cell viability test 1) Transfer 2 brine shrimp into each of the pre-labeled petri dishes containing the solution of NPs and seawater (0, 1:1 & 1:1000) and set a timer for 30 minutes. 2) Transfer a single brine shrimp into the unlabeled petri dish with a drop (~50ul) of seawater and use the dissecting microscope to make observations related to the behavior and morphology of untreated brine shrimp.â¨Make observations in lab notebook of all samples Part D: Preparation of specimens for the toxicity assay 1) 2) 3) 4) 5) Add 10 brine shrimp into each well in the 6 -well plate Make observations in lab notebook Using a dissecting microscope (if needed), count the number of brine shrimp in each well (alive shrimps).â¨Record your data in your notebook using the graphic below as a template. Close the plate and store it for next observation.â¨Observations must be done within 24h-96hâ¨Note: There will be times allocated and a sign up paper for observations Part E: Trypan blue viability test 2 Fall 2017 1) Label 3 glass slides to match the petri dishes prepared in Part C. 2) Transfer 1 brine shrimp from each petri dish onto the corresponding slide. â¨Make observations in your lab notebook 3) Add a drop (~60 Î¼l) of PBS on top of each specimen 4) Add 60 Î¼l of Trypan blue to your shrimp and incubate for 1-2 minutes at room â¨temperature. 5) Wick away excess liquid 6) Add 100 Î¼l PBS to sample and observe (while alive!), looking for blue cells, using the â¨dissecting scope â¨Make observations in your lab notebook 7) Label 3 microfuge tubes to align with each sample. 8) Transfer each brine shrimp from the slide into the corresponding microfuge tube 9) Add 50 Î¼l of PBS to the tube and use the mechanical disruption tool to homogenize the â¨sample for ~2 minutes until a cell suspension has been generated. 10) Load 30-50 Î¼l of your sample onto a new glass slide and add a cover slip. 11)Observe the sample on high power and use the scale below to qualitatively assess cytotoxicityâ 0-no blue 1-some blue 2-fairly blue 3-totally blue.