Exam Questions Chapter 12 Modern Applications Of Microbial

Chapter 12: Modern Applications of Microbial Genetics

= Correct answer

Multiple Choice

1. Which of the following is the definition of biotechnology?
2. The alteration of DNA in a cell
3. The use of living systems to benefit humankind
4. Using computers to synthesize biomolecules
5. Using DNA for solving mathematical problems

Difficulty: Easy

ASM Standard: 26

1. Which of the following is the definition of genetic engineering?
2. The alteration of DNA in a cell
3. The use of living systems to benefit humankind
4. Using computers to synthesize biomolecules
5. Using DNA for solving mathematical problems

Difficulty: Easy

ASM Standard: 15, 19, 26, 31

1. The first set of genes to be introduced into E. coli came from which of the following?
2. Arabidopsis
3. Haemophilus
4. Mycobacterium
5. Xenopus

Difficulty: Easy

ASM Standard: 15, 19

1. Which of the following is a palindrome sequence that could be recognized by a restriction enzyme?

Difficulty: Moderate

ASM Standard: 26, 36

1. Restriction enzymes (endonucleases) are produced by bacteria as a means of protection against which infectious agent?
2. antibiotics
3. antibodies
4. bacteriophages
5. prions

Difficulty: Easy

ASM Standard: 18, 26

1. Which of the following enzymes rejoins two sugar-phosphate backbones of DNA?
2. helicase
3. ligase
4. restriction enzyme
5. reverse transcriptase

Difficulty: Easy

ASM Standard: 26, 36

1. Genes to be cloned into a plasmid are typically inserted in which location?
2. in a location disrupting the antibiotic selective marker
3. oriC (origin of replication)
4. polylinker site
5. randomly

Difficulty: Easy

ASM Standard: 19, 26, 36

1. Which genus of bacterium is naturally competent?
2. Bacillus
3. Escherichia
4. Listeria
5. Yersinia

Difficulty: Moderate

ASM Standard: 11, 19

1. Which of the following regarding pUC19 and blue/white screening is NOT true?
2. Colonies that appear white have the gene of interest inserted into the plasmid pUC19.
3. pUC19 confers antibiotic resistance.
4. The gene of interest being cloned disrupts the lacI gene.
5. X-gal is a chromogenic agent used in blue/white screening to test for β-galactosidase activity.

Difficulty: Moderate

ASM Standard: 19, 26, 36

1. You are in the process of cloning a gene of interest into pUC19 using blue/white screening. You complete your cloning and transform E. coli with your cloned plasmid. You plate your transformation on two media. One medium agar plate contains X-gal only. The second medium agar plate contains X-gal and ampicillin (an antibiotic), in the hope of using blue/white screening. You examine your plates and see a mixture of blue and white colonies. Which of the following colonies should be picked?
2. the blue colonies that are resistant to ampicillin
3. the blue colonies that are sensitive to ampicillin
4. the white colonies that are resistant to ampicillin
5. the white colonies that are sensitive to ampicillin

Difficulty: Moderate

ASM Standard: 15, 19, 26, 28b, 36, 38

1. Which plasmid encodes the genes for its own conjugation?
2. an F-plasmid
3. any plasmid
4. Plasmids cannot be transferred by conjugation.
5. pUC19 only

Difficulty: Easy

ASM Standard: 2, 19, 26, 36

1. Which of the following methods can be used to transfect protoplast plant cells?
2. Agrobacterium tumefaciens
3. electroporation
4. gene gun
5. All these options can be used.

Difficulty: Easy

ASM Standard: 19, 26, 36

1. Which virus is typically used as mechanism to transfect eukaryotic cells?
2. adenovirus
3. bacteriophage
4. influenza virus
5. retrovirus

Difficulty: Easy

ASM Standard: 2, 19, 26, 36

1. For gel electrophoresis, the matrix used for separation of DNA fragments is ________, whereas the matrix used for separation of proteins is ________.
2. agarose; polyacrylamide
3. arabinose; sodium dodecyl sulfate
4. polyacrylamide; agarose
5. sodium dodecyl sulfate; arabinose

Difficulty: Easy

ASM Standard: 36

1. Which charge does a molecule of DNA have?
2. positive
3. neutral
4. negative
5. depends on what the sequence of DNA is

Difficulty: Easy

ASM Standard: 36

1. A ________ blot is used to identify specific sequences of DNA on a membrane, whereas a ________ blot is used to identify specific sequences of RNA on a membrane.
2. Northern; Southern
3. Northern; Western
4. Southern; Northern
5. Western; Eastern

Difficulty: Easy

ASM Standard: 36

1. A two-dimensional PAGE separates molecules according to which two properties?
2. charge at various pHs and hydrophobicity
3. charge at various pHs and size
4. hydrophobicity and presence of disulfide bonds
5. size and hydrophobicity

Difficulty: Moderate

ASM Standard: 36

1. For protein electrophoresis to function, which of the following can be added to the protein to impart a uniform negative charge?
2. ammonium persulfate
3. phosphate
4. polyacrylamide
5. sodium dodecyl sulfate

Difficulty: Easy

ASM Standard: 36

1. You are interested in determining which genes are upregulated in HeLa cells during intracellular infection with a newly discovered pathogen. You prepare a microarray experiment as shown in the diagram below and examine the gene expression of three genes (labeled A through C). Which gene’s expression is not altered during intracellular infection?

1. gene A
2. gene B
3. gene C
4. cannot conclude from this experiment; more information is needed

Difficulty: Difficult

ASM Standard: 28b, 36, 38

1. Which technique can be used to amplify DNA?
2. Genomic libraries
3. PCR
4. Sanger method
5. Southern blot

Difficulty: Easy

ASM Standard: 26, 36

1. Which of the following are the three steps (in order) of PCR?
2. annealing, denaturation, extension
3. denaturation, annealing, extension
4. denaturation, extension, annealing
5. extension, annealing, denaturation

Difficulty: Moderate

ASM Standard: 36

1. Which of the following correctly explains why primers are used in PCR?
2. DNA replication requires a free 3′ hydroxyl group.
3. DNA replication requires a free 3′ phosphate group.
4. DNA replication requires a free 5′ hydroxyl group.
5. DNA replication requires a free 5′ phosphate group.

Difficulty: Easy

ASM Standard: 36

1. The Sanger method/chain termination reaction relies upon using which unique nucleotide?
2. a nucleotide that is missing a 5-carbon sugar
3. a nucleotide that is missing a hydroxyl group
4. a nucleotide that is missing a nitrogenous base
5. a nucleotide that is missing a phosphate group

Difficulty: Moderate

ASM Standard: 36

1. Which of the following is a next-generation sequencing technique in which fragmented DNA has DNA adapters attached, is amplified by PCR, is attached to a bead, and then is placed into a well with sequencing reagents. The flash of light produced by the release of pyrophosphate on addition of a nucleotide is monitored.
2. pyrosequencing
3. Sanger method
4. Southern blot
5. transformation

Difficulty: Moderate

ASM Standard: 36

1. The study of all the DNA of an entire microbial community is known as which of the following?
2. genomics
3. metagenomics
4. metatranscriptomics
5. toxicogenomics

Difficulty: Moderate

ASM Standard: 20, 21, 36

1. In RNAi technology, the dsRNA molecule first combines with which endonuclease that then cleaves it into smaller fragments?
2. DICER
3. Ligase
4. RecA
5. RISC

Difficulty: Hard

ASM Standard: 17, 36

1. Which of the following is/are completely complementary to the target mRNA in RNAi technology?
2. miRNA
3. siRNA
4. both miRNA and siRNA
5. Neither method cleaves mRNA.

Difficulty: Easy

ASM Standard: 17, 36

1. Which disease has been effectively controlled by gene therapy?
2. bacterial meningitis
3. cystic fibrosis
4. ornithine transcarbamylase
5. sickle cell anemia

Difficulty: Moderate

ASM Standard: 19, 26, 31, 36

1. Which agency does not oversee gene therapy?
2. FDA
3. NIH
4. OHRP
5. WHO

Difficulty: Moderate

ASM Standard: 31

1. Genomics would not be an effective method to detect which type of pathogen?
2. bacteria
3. helminths
4. prions
5. viruses

Difficulty: Moderate

ASM Standard: 27

True/False

1. DNA that has been cut with a restriction enzyme, resulting in sticky ends, can be more easily ligated than DNA that has been cut with a restriction enzyme resulting in blunt ends.

Difficulty: Easy

ASM Standard: 19, 26, 36

1. For cloning purposes using transduction, lytic bacteriophages are used.

Difficulty: Easy

ASM Standard: 10, 26

1. Lambda (λ) bacteriophages can be used to make genomic libraries.

Difficulty: Moderate

ASM Standard: 10, 26

1. The uptake of a plasmid by a prokaryotic cell is known as transfection.

Difficulty: Moderate

ASM Standard: 2, 19, 26, 36

1. DNA probes can be used to locate any gene within a cell.

Difficulty: Moderate

ASM Standard: 32, 36

1. Blue/white screening works on the basis of disrupting the lacZ gene in cloning.

Difficulty: Moderate

ASM Standard: 15, 19, 26, 34, 36

1. Reverse transcriptase-PCR amplifies molecules of RNA.

Difficulty: Easy

ASM Standard: 10, 26, 36

1. cDNA is made by reverse transcriptase, which is found in all viruses.

Difficulty: Moderate

ASM Standard: 10, 26, 36

1. Due to concerns about gene transfer, genetically engineered microbes cannot be used to manufacture vaccines.

Difficulty: Easy

ASM Standard: 15, 19, 31

1. RNAi is a natural process used by cells as a means of gene regulation.

Difficulty: Easy

ASM Standard: 17

Matching

1. Match the different steps of molecular cloning to their function.

Answers: A. iv., B. ii., C. v., D. i., E. iii.

Difficulty: Easy

ASM Standard: 19, 26, 36

1. Match the technique to its primary function.

Answers: A. ii., B. iv., C. iii., D. i.

Difficulty: Easy

ASM Standard: 36

1. Match the genetically engineered pharmaceutical to what it can treat.

Answers: A. iii., B. ii., C. i., D. iv.

Difficulty: Moderate

ASM Standard: 26, 31

1. Match the mechanism of horizontal gene transfer to its appropriate description.

Answers: A. iii., B. iv., C. i., D. ii.

Difficulty: Easy

ASM Standard: 2, 26, 36

Fill in the Blank

1. ________ is DNA composed from different organisms.

Difficulty: Easy

ASM Standard: 19, 26, 36

1. ________ are enzymes produced by bacteria that are used in molecular cloning to cut DNA.

Difficulty: Easy

ASM Standard: 26, 36

1. ________ are plasmids that have phage sequences so they can be packaged into bacteriophages.

Difficulty: Moderate

ASM Standard: 10, 18, 26, 36

1. ________ is the technique of using radioactive phosphorous to identify and track sequences of DNA in gel electrophoresis.

Difficulty: Moderate

ASM Standard: 36

1. ________ compares DNA banding patterns of different DNA samples after being incubated with restriction enzymes and subjected to agarose gel electrophoresis.

Difficulty: Easy

ASM Standard: 26, 36

1. ________ is a bacterium that causes crown-gall disease and can be used as a molecular biology tool.

Difficulty: Moderate

ASM Standard: 23, 26, 36

1. ________ is the DNA polymerase that is most commonly used in PCR.

Difficulty: Easy

ASM Standard: 26

1. In proteomics, a ________ is the name given to a protein whose expression is affected by disease.

Difficulty: Easy

ASM Standard: 17, 23, 31, 36

1. A noncoding RNA sequence that is complementary to mRNA is known as ________.

Difficulty: Easy

ASM Standard: 16, 17

1. In gene therapy, genes need to be introduced into ________ to ensure they will be passed on to the next generation.

Difficulty: Easy

ASM Standard: 19, 31, 36

Short Answer

1. Why is it possible for bacteria to produce human proteins such as insulin?

Sample

Difficulty: Difficult

ASM Standard: 16, 19, 26,

1. Why do bacteria produce restriction enzymes (endonucleases)?

Sample

Difficulty: Easy

ASM Standard: 7, 18, 26

1. How can a molecular biologist tell the difference between a chromosome and a commercial plasmid?

Sample

Difficulty: Difficult

ASM Standard: 2, 8, 19, 36

1. What is a reporter gene? Give an example of one.

Sample

Difficulty: Easy

ASM Standard: 17, 19, 26, 36

1. How can bacterial cells become chemically competent?

Sample

Difficulty: Moderate

ASM Standard: 36

1. How does electroporation make bacterial cells competent to receive foreign molecules of DNA?

Sample

Difficulty: Moderate

ASM Standard: 19, 36

1. Why are phages preferred in making a genomic library?

Sample

Difficulty: Moderate

ASM Standard: 26, 36

1. What is the difference between a genomic library and a cDNA library?

Sample

Difficulty: Moderate

ASM Standard: 26, 36

1. Name two molecular techniques one can use to examine gene expression in a bacterial cell.

Sample

Difficulty: Easy

ASM Standard: 17, 26, 36

1. Why is Taq polymerase used in PCR and not E. coli DNA polymerase?

Sample

Difficulty: Easy

ASM Standard: 26, 36

1. Explain how qPCR is quantitative. What is it measuring?

Sample

Difficulty: Moderate

ASM Standard: 36

1. Why is GFP used in the field of proteomics?

Sample

Difficulty: Easy

ASM Standard: 36

1. Which type of disease could be a candidate for treatment using gene therapy?

Sample

Difficulty: Moderate

ASM Standard: 17, 19, 31

1. What are the three steps to PCR?

Sample

Difficulty: Easy

ASM Standard: 26, 36

1. What are the ways transfection can occur?

Sample

Difficulty: Easy

ASM Standard: 19, 36

Brief Essay

Essay Question Rubric

Assume rating/grading scale for the question ranges from 0 to 3 points.

1. Discuss how you can genetically engineer an E. coli cell to produce human insulin. Be sure to include in your answer the use of restriction enzymes, ligase, pUC19, X-gal, selective media, and a method for determining that your transgenic E. coli contains the insulin gene and that it is producing insulin.

Difficulty: Moderate

ASM Standard: 15, 16, 19, 26, 31, 33, 36

1. An outbreak has occurred at a local banquet. You determine that the causative agent was Listeria monocytogenes, which was found in the cold cut meats. You have been asked to further determine from which ribotype the identified L. monocytogenes is. The results of your experiment are shown in the gel diagram below. Answer the questions below the image.

1. What is ribotyping and how is it performed?
2. Which ribotype was found in the cold cut meats?
3. Why do the DNA molecules appear as they do on the gel? Identify where large molecules and small molecules of DNA would be found on this gel.

Difficulty: Moderate

ASM Standard: 5, 26, 28b, 31, 34, 36, 38

1. You wish to amplify a segment of DNA from the genome of the bacterium Haemophilus influenzae. You decide to use PCR. You prepare your PCR using the following reagents and temperatures. What will be the result of each of these? Be sure to explain why these reactions will work or will not work.
2. You add DNA template, DNA primers (forward and reverse), nucleotides, and E. coli DNA polymerase. You set the denaturing temperature for 95 °C, annealing temperature for 50 °C, and the extension temperature for 72 °C. You repeat this for 30 cycles.
3. You add DNA template, nucleotides, and Taq polymerase. You set the denaturing temperature for 95 °C, annealing temperature for 50 °C, and the extension temperature for 72 °C. You repeat this for 30 cycles.
4. You add DNA template, DNA primers (forward and reverse), nucleotides, and Taq polymerase. You set the denaturing temperature for 95 °C, annealing temperature for 70 °C, and the extension temperature for 72 °C. You repeat this for 30 cycles.

Difficulty: Difficult

ASM Standard: 26, 28b, 36

1. Discuss the similarities and differences between miRNA and siRNA.

Sample

Difficulty: Difficult

ASM Standard: 17, 26, 36

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