Deck 11: Genetic Engineering and Biotechnology

Question 1

Which objective would be best to use a Southern blot rather than a Northern blot?

A) Determine if a gene is present in a genome.
B) Discover gene function.
C) Identify regulatory gene-protein interactions.
D) Quantify expression profiles of a gene.

Answer 1

A

Question 2

Inserting a kanamycin resistance cassette into a catabolic operon to confirm the gene is essential in degradation of a particular compound would involve all of the following EXCEPT

A) a reporter gene.
B) ligation.
C) recombination.
D) transformation.

Answer 2

A

Question 3

Which of the following is NOT a common step in creating a genomic library?

A) Fragment DNA into small segments.
B) Hybridize DNA sequences to form inserts of a target size range.
C) Ligate DNA into vectors.
D) Transform the vectors into a host.

Answer 3

B

Question 4

Which statement is TRUE?

A) YACs are more likely than BACs to undergo recombination and rearrangement.
B) BACs are more likely than YACs to undergo recombination and rearrangement.
C) YACs and BACs undergo recombination and rearrangement at about the same rate.
D) It is impossible to state with any certainty whether YACs or BACs are more likely to undergo recombination and rearrangement, because environmental factors play a major role in the probability of one or the other occurring.

Question 5

Detecting a specific protein with an antibody is considered a(n)________ method.

A) selection
B) screening
C) isolation
D) duplication

Question 6

Which of those listed below is LEAST similar in what is measured and concluded?

A) fluorescence in situ hybridization
B) GFP fusion protein
C) Northern blot
D) RT-PCR

Question 7

The genes encoding luciferase,green fluorescent protein (GFP),and β-galactosidase are typically used in cloning as

A) transcription regulators.
B) global control genes.
C) promoter sequences.
D) reporter genes.

Question 8

One of the more formidable obstacles to mammalian gene cloning is the presence of

A) introns.
B) exons.
C) repressors.
D) integrators.

Question 9

At which time period(s)during PCR thermocycling is/are hottest in temperature?

A) during DNA denaturation
B) during primer annealing
C) during primer extension/elongation
D) Both the first and last cycles are hotter in temperature than all other cycles.

Question 10

A polymerase chain reaction (PCR)copies an individual gene segment in vitro with a(n)________ primer(s).

A) individual RNA
B) individual DNA
C) pair of RNA
D) pair of DNA

Question 11

A(n)________ gene is a gene that encodes a protein that is easy to detect and assay.

A) encoder
B) translational
C) reporter
D) recorder

Question 12

To discover a catabolic gene cluster,cloning large sequences of approximately 40 kbp requires the utility of

A) bacterial artificial chromosomes (BACs).
B) cosmids or fosmids.
C) a eukaryotic host to house the large foreign DNA.
D) multiple gene fusions.

Question 13

If a foreign gene is cloned into an expression host,it is important that the host itself

A) not produce the protein being studied.
B) produce the protein in larger amounts than the vector.
C) repress the genetic expression being studied.
D) produce signal proteins to tag the host protein.

Question 14

To estimate the total concentration of a beneficial bacterial species in yogurt,________ would provide the quickest results.

A) fluorescence in situ hybridization
B) qPCR
C) RT-PCR
D) a Southern blot

Question 15

Expression vectors are designed to ensure that ________ can be efficiently ________.

A) mRNA / transcribed
B) DNA / transcribed
C) mRNA / translated
D) DNA / translated

Question 16

What molecular mechanism/feature does site-directed mutagenesis exploit to introduce a mutation at a specific site?

A) flanking complementary bound nucleotides permit non-complementary base pairing
B) methylated nucleotides disrupt DNA polymerase's proofreading
C) nucleotide substitution when one is depleted
D) transposase-induced base pair changes

Question 17

Which of the following is NOT a characteristic of a type II restriction endonuclease?

A) cleavage product can be either blunt or sticky ended but is always the same for an individual enzyme
B) recognizes a specific palindromic site for cleavage
C) recognition site length varies among enzymes but is always the same for an individual enzyme
D) unable to cleave methylated DNA

Question 18

To verify a gene was cloned into a vector successfully,sequencing the vector as well as ________ are commonly performed.

A) agarose gel electrophoresis
B) fluorescence in situ hybridization
C) protein purification
D) northern blots

Question 19

Which of the following sequences would be cleaved by a type II restriction endonuclease?

A) TTGCCGA
AACGGCT
B) GGGGGGG
CCCCCCCC
C) GTAATG
CATTAC
D) GAATTC
CTTAAG

Question 20

What type of vector can replicate and be maintained stably in two (or more)unrelated host organisms?

A) virus
B) expression
C) shuttle
D) integrating

Question 21

Using a host defective in proteases is likely to be necessary when engineering

A) a complete metabolic pathway requiring several different enzymes.
B) overproducing proteins.
C) production of a small protein.
D) transgenic animals with immune systems.

Question 22

The Ti plasmid is best suited for genetically manipulating

A) Agrobacterium spp.
B) fish.
C) plants.
D) viruses.

Question 23

Recognizing pathogens that contain multiple unique proteins which enable the human immune system to recognize just one and mount an effective response has opened the door on development of some vaccines only being

A) attenuated carrier viruses.
B) monovalent.
C) subunit vaccines.
D) purified protein administered.

Question 24

Which of those below is NOT an important consideration when designing a fusion protein construct?

A) Avoid hybridization of the fusion gene in the artificial construct.
B) Reading frame is the same for both the fusion gene and reporter gene.
C) Transcriptional start and stop signals are shared.
D) Translational start and stop signals are shared.

Question 25

If a protein to be overexpressed is toxic to the expression host,it is best to select an expression vector that

A) is compatible with a binary vector able to be regulated.
B) is inducible.
C) has a relatively low copy number per cell.
D) prevents folding of the overexpressed protein into its toxic form.

Question 26

The principle behind a nucleic acid probe design is that the probe itself must contain

A) a key complementary part of the target gene sequence of interest.
B) all of the nucleotide sequence of the gene of interest to conclusively identify the gene.
C) an antibody to specifically bind to the gene of interest.
D) at least three separate complementary regions of the gene of interest.

Question 27

The principle underlying how salmon were genetically engineered to grow faster is the

A) removal of a gene responsible for feeling full after eating.
B) replacement of inducible to constitutive hormone production.
C) resistance to bacterial infections which waste metabolic energy in the salmon to fight off.
D) addition of genes to enhance blood circulation and tissue development.

Question 28

A poorly immunogenic vaccine often suggests the foreign proteins were not properly recognized by the immune system due to a lack of ________ necessary,which can also be engineered to occur with additional molecular manipulations.

A) complex folding
B) methylation
C) glucosylation
D) glycosylation

Question 29

The enzyme that covalently links both strands of a vector and inserted DNA in molecular cloning is

A) DNA ligase.
B) DNA phosphatase.
C) DNA hydrolase.
D) DNA transferase.

Question 30

A shuttle vector is most useful for

A) engineering a complete metabolic pathway.
B) identifying the localization of a protein.
C) knocking out a gene by cassette displacement.
D) making a foreign protein in a mammalian cell.

Question 31

Which of the following terms is used to describe a synthetic DNA fragment?

A) DNA cassette
B) DNA hybrid
C) recombinant DNA
D) artificial chromosome

Question 32

Which construct would be MOST useful in studying translational control?

A) gene fusion
B) operon fusion
C) protein fusion
D) shuttle vector

Question 33

While other types exist,Type II restriction endonucleases are by far the most commonly used enzymes for genetic engineering.

Question 34

What makes eukaryotic transcripts easier to isolate than transcripts in bacteria?

A) Eukaryotic transcripts are not methylated but their genes are often methylated.
B) Larger transcript size in eukaryotes enables easy size-selection methods.
C) mRNA is polyadenylated in eukaryotes.
D) Transcripts are the most abundant RNAs in eukaryotes.

Question 35

After digesting a DNA sequence,a restriction endonuclease can generate

A) blunt ends.
B) overhangs.
C) sticky ends.
D) blunt ends, overhangs, or sticky ends.

Question 36

Polyvalent vaccines using vaccinia virus are highly favored by doctors and physicians but are especially challenging for those who develop them,because

A) coat proteins form a relatively rigid structure and do not allow much space for additional proteins to be expressed.
B) multiple foreign proteins simultaneously synthesized often disrupts each other's activity.
C) vaccinia and most other viruses engineered for vaccines contain only one restriction site for cloning in their genome.
D) virus genetic manipulation uses transfection, which is an inherently inefficient process.

Question 37

Cloning vectors can be distinguished from expression vectors by

A) carrying ori genes for replication of the cloned sequence.
B) having a multiple cloning site (MCS).
C) having a high copy number per cell.
D) lacking a promoter site upstream of the insertion site.

Question 38

Some proteins overexpressed at high levels resulting in the formation of inclusion bodies can abolish the goal of producing large quantities of active protein.What could be done to minimize this issue?

A) Codon optimize the gene.
B) Decrease the number of biobricks in the vector.
C) Simultaneously produce intracellular chaperonins.
D) Switch to an expression host with a larger intracellular volume.

Question 39

Type II restriction endonucleases

A) are heterodimers.
B) natively function to methylate specific nucleotides and prevent foreign DNA from being incorporated into the genome.
C) recognize nucleotide sequences that are palindromic.
D) require ATP energy to cleave dsDNA.

Question 40

Which of the following is NOT an example of synthetic biology?

A) assembling gene sequences together into genome and creating a living organism from it
B) creating a new metabolic pathway that produces a previously unidentified compound
C) developing a novel polyvalent vaccine
D) making Escherichia coli phototrophic

Question 41

DNA ligase mediates the insertion of foreign DNA into a vector,but it will only be able to do so if the inserts and vector have matching sticky or blunt ends.

Question 42

Developing vaccines for humans relies heavily on manipulating and engineering vectors.

Question 43

Green fluorescent protein (GFP)is used for detecting translational activity of a fused protein,whereas lacZ reporters are used to detect transcriptional activity of a fused gene.

Question 44

In principle,a type II restriction endonuclease with an 8-nucleotide recognition sequence should cut 1 in every 4⁸ nucleotide positions.

Question 45

One fundamental technique of genetic engineering includes the ability to cut DNA into random fragments.

Question 46

Modification enzymes typically methylate specific bases within the recognition sequence to prevent digestion of the nucleotide sequence by restriction endonucleases.

Question 47

DNA polymerases from Escherichia coli cannot be used to artificially copy gene sequences with a thermocycler.

Question 48

The lacZ gene is commonly used as a reporter gene,because its substrate lactose is well known and easily measured.

Question 49

Regardless of the DNA polymerase used in PCR,such as Taq or Pfu,they all have an inherent inability to perfectly copy the template strand,which means the polymerases themselves occasionally make mutations in the sequences they copy.

Question 50

High expression levels of a eukaryotic gene in a bacterium such as Escherichia coli cannot be accomplished due to the presence of introns.

Question 51

Artificially synthesizing DNA strands (e.g.,oligonucleotide primers)involves the careful attachment of one nucleotide at a time to an immobilized sequence.

Question 52

Genomic libraries enable the discovery of individual gene(s)involved in a particular function of interest with cloning vectors in an expression host,such as Escherichia coli.

Question 53

Due to well developed molecular tools and careful screening designs,functional genes can be isolated directly by isolation from the environment rather than cultivating the diverse species in a microbial community.

Question 54

Engineering a metabolic pathway enables a researcher to use different genes from unrelated organisms.

Question 55

The key steps in cloning a foreign gene into a vector,regardless of the application,involve isolating the insert fragment,ligating the insert into a vector,and transforming it into a host.

Question 56

Although various codons often code for the same amino acid,it is important to choose the codon preferred by the expression host itself.

Question 57

Strong promoters used for genetic manipulation are usually regulated by specific molecules.

Question 58

One method to circumvent issues with introns when expression eukaryotic gene is a bacterium is to simply clone the mature transcript.

Question 59

One problem with both BACs and YACs is that genetic regions of these chromosomes cannot be subcloned.

Question 60

One important advantage of eukaryotic cells as hosts for cloning vectors is that they already possess the complex RNA and posttranslational processing systems required for the production of eukaryotic proteins.

Question 61

When trying to make a mammalian protein in a bacterium,expression vectors are often used.Using your knowledge of prokaryotic and eukaryotic genetics,explain why expression vectors are so important.

Question 62

How can foreign DNA of greater than 300 kbp be inserted and stably maintained in BAC vectors?

Question 63

What is necessary for a YAC to function as a normal eukaryotic chromosome?

Question 64

Explain why DNA fragments migrate toward the positive electrode and why some fragments migrate more rapidly than others during gel electrophoresis.How does this electronegativity influence observations and conclusions drawn from DNA migrated through an agarose gel?

Question 65

If vaccinia viruses were not both immunogenic and relatively benign,they would likely not be a favored vehicle for vaccinations.

Question 66

Compare how the process of cloning differs when a vector with sticky ends is used and when a vector with blunt ends is used.

Question 67

Describe the methodology employed when antibodies are used to detect proteins.

Question 68

Explain the process of site-directed mutagenesis,and discuss some applications of this technique.

Question 69

Describe three characteristics of an ideal cloning host for genetic engineering.

Question 70

Describe a method developed by molecular biologists to easily observe the success of a genetic engineering procedure involving the ligating of a gene of interest into a plasmid.

Question 71

Create an experiment where you would use both a Southern blot and a Northern blot to determine if a newly isolated bacterium contains and actively uses the same catabolic pathway as previously demonstrated.What is the value in doing the Southern blot first? With results from these experiments,how would you conclude if the same pathway is active or not?

Question 72

When is it appropriate to use an artificial chromosome vector? Describe a specific example.

Question 73

Defend why codon usage must be considered when cloning a gene into another organism.

Question 74

Describe the three main steps to clone a gene into an organism.

Question 75

If you wanted to discover a novel biosynthetic pathway for an antimicrobial compound and were given a handful of soil,describe the key experimental steps you would need to take to accomplish the task and explain why alternative procedures were not performed.

Question 76

Describe the usefulness of blue-white screening,also called α-complementation,in cloning vectors such as pUC19.Include in your answer the terms polylinker,DNA ligase,lacZ gene,insertional inactivation,Xgal,and β-galactosidase.

Question 77

Defend how type II restriction endonucleases,which do not cleave random sites,can be a part of an approach to random cloning library.

Question 78

Explain how T7 promoters regulate transcription and why cloning vectors contain them.

Question 79

Describe the procedure used for colony hybridization and why it is useful.

Question 80

Explain how a DNA library is created and how a particular vector is selected.