Deck 11: Genetic Engineering

Question 1

Which construct would be MOST useful in studying translational control?

A) operon fusion
B) gene fusion
C) protein fusion
D) all of the above

Answer 1

C

Question 2

M13 is an ssDNA filamentous phage used for

A) hybridization.
B) creating gene fusions.
C) assembling vectors.
D) site- directed mutagenesis.

Answer 2

D

Question 3

Which is the organism of choice for most molecular cloning?

A) Escherichia coli
B) Bacillus subtilis
C) Vibrio natriegens
D) Saccharomyces cerevisiae

Answer 3

A

Question 4

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

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

Question 5

A vector that can be used to clone a desired gene that contains regulatory sequences so the expression of the gene can be manipulated is called a(n) vector.

A) stability
B) expression
C) endogenous
D) regulatory

Question 6

mRNA transcripts are converted into cDNA by

A) a restriction endonuclease.
B) reverse transcriptase.
C) DNA polymerase.
D) RNA polymerase.

Question 7

Molecular cloning includes

A) locating the gene of interest.
B) placing the gene of interest into a convenient vector.
C) obtaining and purifying a copy of the gene of interest.
D) all of the above.

Question 8

To make an adequate library for sequencing prokaryotic genes, a vector should hold inserts in the range of

A) 10- 50 kbp.
B) 0.5- l kbp.
C) 50- 100 kbp.
D) 2- 10 kbp.

Question 9

An illustration that indicates all regions where a particular restriction endonuclease cleaves a DNA sequence is called a(n) map.

A) shotgun
B) reporter
C) restriction
D) cleavage

Question 10

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

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

Question 11

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

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

Question 12

Which of the following genomes have almost no introns?

A) those belonging to microbial eukaryotes
B) those belonging to prokaryotes
C) those belonging to plants
D) those belonging to animals

Question 13

The principle behind a nucleic acid probe is that the probe itself contains

A) at least three separate sequences of the gene of interest to minimize false positives.
B) an antibody to specifically bind to the gene of interest.
C) a key part of the base sequence of the gene of interest.
D) all of the nucleotide sequence of the gene of interest to conclusively identify the gene.

Question 14

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

A) operon
B) DNA cassette
C) DNA hybrid
D) DNA probe

Question 15

By definition, genetic engineering techniques are performed

A) in toto.
B) in vivo.
C) in situ.
D) in vitro.

Question 16

Bacterial artificial chromosomes (BACs) can be constructed using the

A) R plasmid.
B) M plasmid.
C) plasmid.
D) M13 phage.

Question 17

Which of the following sequences is a palindrome?

A) GAATTC CTTAAG

Question 18

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

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

Question 19

Which statement is CORRECT about genomic analysis?

A) It does not matter whether sequencing or cloning occurs first.
B) Sequencing of DNA precedes cloning of fragments.
C) Cloning of fragments precedes sequencing of DNA.
D) Sequencing of DNA and cloning of fragments are done simultaneously.

Question 20

The class of restriction endonucleases that cleave the DNA within their recognition sequences is known as

A) type III.
B) type II.
C) type I.
D) All restriction endonucleases cut within their recognition sequence.

Question 21

Two problems of using Escherichia coli as a cloning host are and .

Question 22

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 transferase.
D) DNA hydrolase.

Question 23

After digesting a DNA sequence, a restriction endonuclease can generate

A) overhangs.
B) blunt ends.
C) sticky ends.
D) all of the above.

Question 24

An integrating vector is a specific type of plasmid that is used for .

Question 25

The is a naturally occurring plasmid within yeast cells, and development of cloning yeast vectors is often based on this particular plasmid.

Question 26

Type II restriction endonucleases

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

Question 27

Retroviruses can be used to introduce foreign genes into mammalian cells, because
.

Question 28

After DNA has been electrophoresed through a(n) , a stain such as ethidium bromide is used to visualize different DNA sequence .

Question 29

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

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

Question 30

Nucleic acid hybridization is widely used for segments of DNA.

A) identifying
B) characterizing
C) detecting
D) all of the above.

Question 31

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

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

Question 32

Most vectors are derived from

A) mRNA.
B) plasmids or viruses.
C) bacterial genomes.
D) human chromosomes.

Question 33

Restriction enzymes are widespread among , where they serve a protective function.

Question 34

Which statement is TRUE?

A) YACs are more likely than BACs to undergo recombination and rearrangement.
B) YACs and BACs undergo recombination and rearrangement at about the same rate.
C) BACs are more likely than YACs to undergo recombination and rearrangement.
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 35

A(n) mutation is one in which a DNA cassette is inserted at a restriction site in a cloned gene; transformed cells lose the function of the gene into which the cassette is inserted.

Question 36

Expression vectors used for cloning bacterial genes into eukaryotic cells must contain a(n)
that functions efficiently in the host and one that is correctly positioned to drive transcription of the cloned gene.

Question 37

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

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

Question 38

replicate in a host such as a plasmid but are also able to be packaged into phage virions. These vectors can carry large inserts of up to 50 kbp, so they do not efficiently transform; instead they enter into a host through .

Question 39

Which of the following procedures was named after its developer?

A) Northern blot
B) Western blot
C) Southern blot
D) Eastern blot

Question 40

Hybridization studies require the use of _, which are segments of single- stranded nucleic acids whose identity is already known.

Question 41

Recombinant vaccines have been developed using .

Question 42

The host for a BAC is typically a mutant strain of Escherichia coli that is missing and is defective in _.

Question 43

It is usually desirable to design a vector that transcribes cloned genes at high levels at all times.

Question 44

One limitation to designing an antibody for detecting a particular protein is that the antibody must be for the protein of interest.

Question 45

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 46

Although a gene library may contain tens of thousands of clones, finding the clone of interest is the biggest challenge.

Question 47

A(n) uses a DNA or RNA probe to detect the presence of a particular DNA sequence. A(n) uses a DNA or RNA probe that is used to detect the presence of a particular RNA sequence.

Question 48

A major disadvantage of using Bacillus subtilis as a cloning host is that over several generations of subculturing, .

Question 49

Artificial chromosomes are vectors capable of carrying .

Question 50

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

Question 51

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

Question 52

If the gene in question is not expressed and no assay or antibody is available to detect the gene product, the most common procedure used for identification is a(n) probe.

Question 53

One method of avoiding false positive results during hybridization reactions is to adjust the of the hybridization, such that complementarity between the sequence of the gene of interest and the probe must be nearly exact.

Question 54

Banding patterns generated from restriction analyses of either whole chromosomes or specific genes from a series of organisms can yield an indication of their genetic relationships.

Question 55

Codon bias offers clues about reading frame functionality because most organisms show preferences among .

Question 56

Lambda is a particularly useful cloning vector because and _ .

Question 57

A cloning vector that can stably replicate in two unrelated host organisms is known as a(n)
.

Question 58

The process by which a fragment of DNA is placed into a vector where it can be replicated is called .

Question 59

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

Question 60

Genetic engineering includes techniques that result in genetically modified organisms.

Question 61

Explain the process of microinjection.

Question 62

Site- directed mutagenesis is an in vitro method that removes mutations at specific locations in a sequence.

Question 63

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

Question 64

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

Question 65

Often organisms used as hosts for genetic engineering must have specific genotypes to be effective.

Question 66

Cloning vectors are small, independently replicating genetic elements used to replicate cloned genes.

Question 67

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 68

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

Question 69

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

Question 70

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

Question 71

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

Question 72

Explain how foreign DNA of greater than 300 kbp can be inserted and stably maintained in BAC vectors.

Question 73

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

Question 74

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

Question 75

Viral vectors can be used for molecular cloning, but plasmid vectors cannot.

Question 76

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

Question 77

Explain how a DNA library is created.

Question 78

Compare and contrast operon and protein fusions.

Question 79

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

Question 80

Explain the function and usefulness of a polylinker within a vector.