Concept: Transformation and Restriction Endonucleases6m
Concept: Recombinant Plasmids5m
Concept: Transformation and Antibiotic Resistance6m
During the procedure to form recombinant DNA molecules, the purpose of the cell transformation step is to
A. use the transformed cells to combine vector and source DNA in order to form recombinant molecules.
B. use the transformed cells as a source of DNA ligase to form recombinant molecules.
C. use the transformed cells to cut the DNA at specific sites to generate fragments.
D. all the above
E. None of the above
Name the restriction endonuclease that cuts at the following sites. Indicate whether it cuts in a blunt or staggered manner.
What is recombinant DNA technology? What are the safety issues related to recombinant DNA technology?
You digest DNA from the frog Xenopus laevis with HindIII that produces sticky DNA ends. You can ligate your DNA to : (check all that apply)
A. HindIII sticky DNA ends from Xenopus laevis
B. EcoRI sticky DNA ends from Xenopus laevis
C. blunt DNA ends from bacteria
D. HindIII sticky DNA ends from bacteria
Which ofthe following is a palindromic restriction site?
A. 5' - ACGGCA - 3'
B. 5' - TGCGCC - 3'
C. 5' - AAGGTT - 3'
D. 5' - ACGCGT - 3'
E. 5' - GCAGCA - 3'
Jason is setting up a reaction with the restriction endonuclease EcoRI. The concentration of the EcoRI enzyme solution is 20,000 units/mL. Jason mixes the following in a test tube:
4 uL dsDNA, 14 uL buffer, 2 uL EcoRI
What is the final concentration of EcoRI enzyme in Jason's reaction?
A. 2,222 units/mL
B. 2,857 units/mL
C. 2,000 units/mL
D. 200,000 units/mL
How is a gene for a particular protein inserted into a plasmid?
What is a restriction enzyme? What role do they play in biotechnology experiments?
What is Recombinant DNA?
Restriction enzymes cut DNA at random sites.
"Sticky ends" are
a. single-stranded DNA sequences that are generated by staggered cuts.
b. double-stranded DNA sequences that are generated by staggered cuts.
c. single-stranded DNA sequences that are generated by blunt cuts.
d. double-stranded DNA sequences that are generated by blunt cuts.
The function of the enzyme ligase is to
a. add new DNA to the plasmid.
b. remove DNA from the plasmid.
c. form covalent bonds between cloned gene and plasmid.
d. repair damaged DNA.
e. purify the plasmid DNA.
Arrange the following events in the proper sequence for gene cloning.
1 = Incorporate gene into bacterial plasmid
2 = Isolate DNA from organism containing desired gene
3 = Incorporate cloned gene into bacterial cells
4 = Fragment DNA with restriction enzyme
a. 1, 2, 3, 4
b. 2, 1, 4, 3
c. 2, 3, 4, 1
d. 2, 4, 1, 3
e. 2, 4, 3, 1
Restriction enzymes cut only at specific sites and therefore are not useful for genetic engineering.
Human DNA cut with restriction enzyme A can be joined to
a. human DNA cut with restriction enzyme B.
b. human DNA that is uncut.
c. bacterial DNA cut with restriction enzyme A.
d. bacterial DNA that is uncut.
e. none of the above
The single-stranded ends of DNA molecules can be joined together by
a. restriction endonucleases.
b. DNA ligase.
c. DNA polymerase.
The single-stranded but complementary ends of the two DNA molecules cleaved by the restriction enzyme _____.
a. are called "sticky ends"
b. can bind by complementary base pairing
c. facilitate the insertion of foreign DNA into vector DNA
d. All of the above
The foreign DNA is sealed into the opening created by the restriction enzyme by _____.
a. DNA polymerase
b. DNA repair enzymes
c. DNA ligase
The plasmid DNA is cleaved by _____.
a. DNA ligase
b. DNA polymerase
c. a restriction enzyme
Small accessory rings of DNA are called _____.
a. recombinant DNA
The process used to rapidly produce DNA sequences of interest is DNA sequencing.