4

1. Restriction enzymes were discovered by

A. B

oyer and Cohen.
B. L

inn and Arber.
C. Watson and Crick.

D. Gurdon and Linn.

E. C
ohen and Gurdon.

2. A new enzyme was isolated and found to have a recognition sequence of 5 base pairs. How often
will it cut in a genome?

A. 6

4
B. 4

096
C. 8192

D. 1024

E. 3
124

3. The complementary sequence of one of the following is a palindrome. Which one?

A. G

AATTC
B. C

TGACT
C. ATCCTC

D. GGCCAA

E. C
CTTTC

4. Which of the following enzymes will produce blunt-ended cuts in DNA?

A. E

coRI
B. H

indIII
C. SmaI

D. BamHI

E. P
stI


5. Which of the following enzymes cannot catalyze the formation of a phosphodiester bond?

A. e

ndonuclease
B. R

NA polymerase
C. DNA polymerase

D. ligase

E. n
one of the choices are correct.

6. Select the component that is most critical to the function of a plasmid.

A. t etracycline resistance

B. a

mpicillin resistance
C. EcoRI site

D. origin of replication

E. l igase

7. A molecular biologist is able to select recombinant colonies from a bacterial culture plate based
on the color of the colonies. Which of the following is a correct statement?

A. C

olonies that contain recombinant plasmids are red.
olonies that contain recombinant plasmids are blue due to the disruption of the lacZ gene the
B. C

insert.
C. Colonies that contain recombinants are clear/white due to the disruption of the lacZ gene by

the insert.
D. Colonies are blue due to the disruption of the ampicillin gene.

E. A
ll of the choices are correct.

8. One approach to prevent religation of the ends of a restricted plasmid DNA is to

A. s tore the plasmid in DNase.

B. s tore the plasmid in RNAase.

C. treat the plasmid with alkaline phosphatase to remove phosphates from the 'sticky ends'

D. treat the plasmid with a protease to remove all proteins.

E. t reat the plasmid with BamHI to create gaps.

9. A recombinant plasmid was constructed with several restriction enzyme sites and an ampicillin
resistance marker. This recombinant was used to transform a host bacteria that also has the
ampicillin resistant gene. Which of the following statements is true regarding this experiment?

A. A

ll transformed bacteria will grow on a plate containing ampicillin.
B. A

ll untransformed bacteria will grow on a plate containing ampicillin.
C. It will be difficult to screen for a colony containing a plasmid.

D. A bacterial host that is ampicillin-sensitive should be used for the screening.

E. A
ll of the choices are true.

10. Which of the following is true regarding alpha complementation?

A. T
he plasmid encodes the amino terminal of β-galactosidase while the host bacterium encodes

the carboxyl portion .
he plasmid encodes both amino and carboxyl terminals of the lacZ region.
B. T

C. The complete β-galactosidase enzyme is made by the host.

D. When the alpha and beta regions fuse they form an inactive peptide.

11. A bacterial sample was contaminated with an unknown preparation of vector DNA. In order to
identify the vector, the bacteria was streaked on a plate and incubated overnight. Examination of
the plate revealed at least 120 clear plaques. Which of the following is a plausible conclusion?

A. T

he vector could be a plasmid.
B. T

he vector could be a phage.
C. The vector is a Ti.

D. The vector is puc18.

E. T
he vector is a cosmid.

12. Which of the following vectors is useful in producing single-stranded DNA?

A. p

uc18
B. λ phage

C. M13

D. BAC

E. Y AC


13. What is the RACE technique?

A. s creening a genomic library

B. e

xtending incomplete cDNA sequences
C. generating polynucleotide probes

D. rapid amplification of genomic DNA

E. s creening a genomic library and generating polynucleotide probes

14. The correct 10-base forward PCR primer starting at position 5 on the following sequence would
be 5′ -ATGCCGATGTAGGGCGGGATGGAGAGATAGAGAGAGTCACAAT-3′

A. 5

-GGGATGGCAT
B. 5

′-ATGCCGATGT
C. 5′-CGATGTAGGG

D. 5′-GAGTCACAAT

E. 5
′-GCTACATCCC

15. Which of the following is ideal for screening a protein expression library?

A. l abeled antibodies

B. 3

′ RACE
C. 5′ RACE

D. RT-PCR

E. R eal-Time PCR

16. Which of the following vectors is the best choice for the expression of eukaryotic proteins?

A. M

13
B. b

aculovirus
C. phagemid

D. pUC18

E. p
UC19


17. A disadvantage of using a prokaryotic expression system for eukaryotic proteins is that the
proteins are

A. h

ighly phosphorylated after translation.
B. i mproperly folded.

C. highly soluble.

D. heavily glycosylated.

E. o ver expressed.

18. The "Flavr Savr" tomato is generated by

A. t he introduction of plasmids into tomatoes.

B. i ntroducing the antisense copy of the gene that contributes to ripening.

C. the inhibition of transcription of most genes in the tomato.

D. the introduction of the sense copy of the gene that contributes to ripening.

E. t he introduction of plasmids into tomatoes and the introduction of the sense copy of the gene
that contributes to ripening are correct

19. How many enzymes should be used to cut the plasmid DNA and the insert DNA if there is a
desire to ligate the insert within the vector in a specific orientation?

A. 1


B. 2


C. 3

D. 4

E. 5

20. In constructing a cDNA library, which of the following would you use to generate rare restriction
sites on the cDNA strands?

A. r estriction enzymes

B. P

CR with multiple cycles
C. BamHI restriction followed by ligation

D. ligate cDNA to specific linkers

E. 5 ′ RACE


21. In the construction of an expression vector, which of the following would you include in order to
stimulate a high level of RNA synthesis?

A. P

3
B. T

7 phage promoter
C. Ampr gene

D. His region

E. G
FP

22. A group of identical cells is called a __________.

________________________________________

23. An enzyme that recognizes different sites in an identical sequence is called a ____________.

________________________________________

24. The use of high voltage to drive DNA into cells is called _____________.

________________________________________

25. A fusion protein can easily be isolated by ____________________ chromatography.

________________________________________

26. HindIII is so named because it was isolated from Haemophilus influenzae bacterium.

True False

27. Bacteria use the restriction-modification system to synthesize new DNA molecules.

True False

28. A fragment restricted with EcoRI enzyme can be used for ligation into a plasmid that was
restricted with BamHI because both the insert and the plasmid contain sticky ends.

True False

29. In a replacement vector, a portion of the DNA is removed to accommodate the fragment to be
inserted.

True False

30. Polynucleotide probes can be used to screen a genomic library for specific genomic sequences.

True False

 4 Key

1. Restriction enzymes were discovered by

A. Boyer and Cohen.

B. Linn and Arber.

C. Watson and Crick.

D. Gurdon and Linn.

E. Cohen and Gurdon.

Bloom's Level: 1. Remember

Section: 04.01
Weaver - Chapter 04 #1


2. A new enzyme was isolated and found to have a recognition sequence of 5 base pairs. How
often will it cut in a genome?

A. 64

B. 4096

C. 8192

D. 1024

E. 3124

Bloom's Level: 2. Understand

Section: 04.01
Weaver - Chapter 04 #2


3. The complementary sequence of one of the following is a palindrome. Which one?

A. GAATTC

B. CTGACT

C. ATCCTC

D. GGCCAA

E. CCTTTC

Bloom's Level: 2. Understand

Section: 04.01
Weaver - Chapter 04 #3



4. Which of the following enzymes will produce blunt-ended cuts in DNA?

A. EcoRI

B. HindIII

C. SmaI

D. BamHI

E. PstI

Bloom's Level: 1. Remember

Section: 04.01
Weaver - Chapter 04 #4


5. Which of the following enzymes cannot catalyze the formation of a phosphodiester bond?

A. endonuclease

B. RNA polymerase

C. DNA polymerase

D. ligase

E. none of the choices are correct.

Bloom's Level: 2. Understand

Section: 04.01
Weaver - Chapter 04 #5


6. Select the component that is most critical to the function of a plasmid.

A. tetracycline resistance

B. ampicillin resistance

C. EcoRI site

D. origin of replication

E. ligase

Bloom's Level: 2. Understand

Section: 04.01
Weaver - Chapter 04 #6



7. A molecular biologist is able to select recombinant colonies from a bacterial culture plate
based on the color of the colonies. Which of the following is a correct statement?

A. Colonies that contain recombinant plasmids are red.

B. Colonies that contain recombinant plasmids are blue due to the disruption of the lacZ gene

the insert.
C. Colonies that contain recombinants are clear/white due to the disruption of the lacZ gene by

the insert.
D. Colonies are blue due to the disruption of the ampicillin gene.

E. All of the choices are correct.

Bloom's Level: 2. Understand

Section: 04.01
Weaver - Chapter 04 #7


8. One approach to prevent religation of the ends of a restricted plasmid DNA is to

A. store the plasmid in DNase.

B. store the plasmid in RNAase.

C. treat the plasmid with alkaline phosphatase to remove phosphates from the 'sticky ends'

D. treat the plasmid with a protease to remove all proteins.

E. treat the plasmid with BamHI to create gaps.

Bloom's Level: 3. Apply

Section: 04.01
Weaver - Chapter 04 #8


9. A recombinant plasmid was constructed with several restriction enzyme sites and an ampicillin
resistance marker. This recombinant was used to transform a host bacteria that also has the
ampicillin resistant gene. Which of the following statements is true regarding this experiment?

A. All transformed bacteria will grow on a plate containing ampicillin.

B. All untransformed bacteria will grow on a plate containing ampicillin.

C. It will be difficult to screen for a colony containing a plasmid.

D. A bacterial host that is ampicillin-sensitive should be used for the screening.

E. All of the choices are true.

Bloom's Level: 5. Evaluate

Section: 04.01
Weaver - Chapter 04 #9



10. Which of the following is true regarding alpha complementation?

A. The plasmid encodes the amino terminal of β-galactosidase while the host bacterium

encodes the carboxyl portion .
B. The plasmid encodes both amino and carboxyl terminals of the lacZ region.

C. The complete β-galactosidase enzyme is made by the host.

D. When the alpha and beta regions fuse they form an inactive peptide.

Bloom's Level: 1. Remember

Section: 04.01
Weaver - Chapter 04 #10


11. A bacterial sample was contaminated with an unknown preparation of vector DNA. In order to
identify the vector, the bacteria was streaked on a plate and incubated overnight. Examination
of the plate revealed at least 120 clear plaques. Which of the following is a plausible
conclusion?

A. The vector could be a plasmid.

B. The vector could be a phage.

C. The vector is a Ti.

D. The vector is puc18.

E. The vector is a cosmid.

Bloom's Level: 2. Understand

Section: 04.01
Weaver - Chapter 04 #11


12. Which of the following vectors is useful in producing single-stranded DNA?

A. puc18

B. λ phage

C. M13

D. BAC

E. YAC

Bloom's Level: 2. Understand

Section: 04.01
Weaver - Chapter 04 #12



13. What is the RACE technique?

A. screening a genomic library

B. extending incomplete cDNA sequences

C. generating polynucleotide probes

D. rapid amplification of genomic DNA

E. screening a genomic library and generating polynucleotide probes

Bloom's Level: 2. Understand

Section: 04.01
Weaver - Chapter 04 #13


14. The correct 10-base forward PCR primer starting at position 5 on the following sequence
would be 5′ -ATGCCGATGTAGGGCGGGATGGAGAGATAGAGAGAGTCACAAT-3′

A. 5-GGGATGGCAT

B. 5′-ATGCCGATGT

C. 5′-CGATGTAGGG

D. 5′-GAGTCACAAT

E. 5′-GCTACATCCC

Bloom's Level: 5. Evaluate

Section: 04.01
Weaver - Chapter 04 #14


15. Which of the following is ideal for screening a protein expression library?

A. labeled antibodies

B. 3′ RACE

C. 5′ RACE

D. RT-PCR

E. Real-Time PCR

Bloom's Level: 2. Understand

Section: 04.01
Weaver - Chapter 04 #15



16. Which of the following vectors is the best choice for the expression of eukaryotic proteins?

A. M13

B. baculovirus

C. phagemid

D. pUC18

E. pUC19

Bloom's Level: 2. Understand

Section: 04.03
Weaver - Chapter 04 #16


17. A disadvantage of using a prokaryotic expression system for eukaryotic proteins is that the
proteins are

A. highly phosphorylated after translation.

B. improperly folded.

C. highly soluble.

D. heavily glycosylated.

E. over expressed.

Bloom's Level: 2. Understand

Section: 04.03
Weaver - Chapter 04 #17


18. The "Flavr Savr" tomato is generated by

A. the introduction of plasmids into tomatoes.

B. introducing the antisense copy of the gene that contributes to ripening.

C. the inhibition of transcription of most genes in the tomato.

D. the introduction of the sense copy of the gene that contributes to ripening.

E. the introduction of plasmids into tomatoes and the introduction of the sense copy of the
gene that contributes to ripening are correct

Bloom's Level: 1. Remember

Section: 04.03
Weaver - Chapter 04 #18



19. How many enzymes should be used to cut the plasmid DNA and the insert DNA if there is a
desire to ligate the insert within the vector in a specific orientation?

A. 1

B. 2

C. 3

D. 4

E. 5

Bloom's Level: 2. Understand

Section: 04.01
Weaver - Chapter 04 #19


20. In constructing a cDNA library, which of the following would you use to generate rare
restriction sites on the cDNA strands?

A. restriction enzymes

B. PCR with multiple cycles

C. BamHI restriction followed by ligation

D. ligate cDNA to specific linkers

E. 5′ RACE

Bloom's Level: 6. Create

Section: 04.01
Weaver - Chapter 04 #20


21. In the construction of an expression vector, which of the following would you include in order
to stimulate a high level of RNA synthesis?

A. P3

B. T7 phage promoter

C. Ampr gene

D. His region

E. GFP

Bloom's Level: 2. Understand

Section: 04.03
Weaver - Chapter 04 #21


22. A group of identical cells is called a __________.

clone

Bloom's Level: 1. Remember
Section: 04.01
Weaver - Chapter 04 #22



23. An enzyme that recognizes different sites in an identical sequence is called a ____________.

heteroschizomer

Bloom's Level: 2. Understand
Section: 04.01
Weaver - Chapter 04 #23


24. The use of high voltage to drive DNA into cells is called _____________.

electroporation

Bloom's Level: 2. Understand
Section: 04.03
Weaver - Chapter 04 #24


25. A fusion protein can easily be isolated by ____________________ chromatography.

affinity

Bloom's Level: 2. Understand
Section: 04.03
Weaver - Chapter 04 #25


26. HindIII is so named because it was isolated from Haemophilus influenzae bacterium.

TRUE

Bloom's Level: 2. Understand
Section: 04.01
Weaver - Chapter 04 #26


27. Bacteria use the restriction-modification system to synthesize new DNA molecules.

FALSE

Bloom's Level: 2. Understand
Section: 04.01
Weaver - Chapter 04 #27


28. A fragment restricted with EcoRI enzyme can be used for ligation into a plasmid that was
restricted with BamHI because both the insert and the plasmid contain sticky ends.

FALSE

Bloom's Level: 2. Understand
Section: 04.01
Weaver - Chapter 04 #28



29. In a replacement vector, a portion of the DNA is removed to accommodate the fragment to be
inserted.

TRUE

Bloom's Level: 2. Understand
Section: 04.01
Weaver - Chapter 04 #29


30. Polynucleotide probes can be used to screen a genomic library for specific genomic
sequences.

TRUE

Bloom's Level: 2. Understand
Section: 04.03
Weaver - Chapter 04 #30

 4 Summary

Category # of Questions
Bloom's Level: 1. Remember 5
Bloom's Level: 2. Understand 21
Bloom's Level: 3. Apply 1
Bloom's Level: 5. Evaluate 2
Bloom's Level: 6. Create 1
Section: 04.01 23
Section: 04.03 7
Weaver - Chapter 04 30