Chapter 20 Flashcards
1) Assume that you are trying to insert a gene into a plasmid. Someone gives you a preparation of genomic DNA that has been cut with restriction enzyme X. The gene you wish to insert has sites on both ends for cutting by restriction enzyme Y. You have a plasmid with a single site for Y, but not for X. Your strategy should be to
A) insert the fragments cut with restriction enzyme X directly into the plasmid without cutting the plasmid.
B) cut the plasmid with restriction enzyme X and insert the fragments cut with restriction enzyme Y into the plasmid.
C) cut the DNA again with restriction enzyme Y and insert these fragments into the plasmid cut with the same enzyme.
D) cut the plasmid twice with restriction enzyme Y and ligate the two fragments onto the ends of the DNA fragments cut with restriction enzyme X.
E) cut the plasmid with restriction enzyme X and then insert the gene into the plasmid.
C) cut the DNA again with restriction enzyme Y and insert these fragments into the plasmid cut with the same enzyme.
2) How does a bacterial cell protect its own DNA from restriction enzymes?
A) by adding methyl groups to adenines and cytosines
B) by using DNA ligase to seal the bacterial DNA into a closed circle
C) by adding histones to protect the double-stranded DNA
D) by forming “sticky ends” of bacterial DNA to prevent the enzyme from attaching
E) by reinforcing the bacterial DNA structure with covalent phosphodiester bonds
A) by adding methyl groups to adenines and cytosines
3) What is the most logical sequence of steps for splicing foreign DNA into a plasmid and inserting the plasmid into a bacterium?
I. Transform bacteria with a recombinant DNA molecule.
II. Cut the plasmid DNA using restriction enzymes.
III. Extract plasmid DNA from bacterial cells.
IV. Hydrogen-bond the plasmid DNA to nonplasmid DNA fragments.
V. Use ligase to seal plasmid DNA to nonplasmid DNA.
A) I, II, IV, III, V B) II, III, V, IV, I C) III, II, IV, V, I D) III, IV, V, I, II E) IV, V, I, II, III
C) III, II, IV, V, I
4) A principal problem with inserting an unmodified mammalian gene into a BAC, and then getting that gene expressed in bacteria, is that
A) prokaryotes use a different genetic code from that of eukaryotes.
B) bacteria translate polycistronic messages only.
C) bacteria cannot remove eukaryotic introns.
D) bacterial RNA polymerase cannot make RNA complementary to mammalian DNA.
E) bacterial DNA is not found in a membrane-bounded nucleus and is therefore incompatible with mammalian DNA.
C) bacteria cannot remove eukaryotic introns.
5) A gene that contains introns can be made shorter (but remain functional) for genetic engineering purposes by using
A) RNA polymerase to transcribe the gene.
B) a restriction enzyme to cut the gene into shorter pieces.
C) reverse transcriptase to reconstruct the gene from its mRNA.
D) DNA polymerase to reconstruct the gene from its polypeptide product.
E) DNA ligase to put together fragments of the DNA that code for a particular polypeptide.
C) reverse transcriptase to reconstruct the gene from its mRNA.
Why are yeast cells frequently used as hosts for cloning?
A) They easily form colonies.
B) They can remove exons from mRNA.
C) They do not have plasmids.
D) They are eukaryotic cells.
E) Only yeast cells allow the gene to be cloned.
D) They are eukaryotic cells.
The DNA fragments making up a genomic library are generally contained in A) BACs. B) recombinant viral RNA. C) individual wells. D) DNA-RNA hybrids. E) radioactive eukaryotic cells.
A) BACs.
Yeast artificial chromosomes contain which of the following elements?
A) centromeres only
B) telomeres only
C) origin of replication only
D) centromeres and telomeres only
E) centromeres, telomeres, and an origin of replication
E) centromeres, telomeres, and an origin of replication
Which of the following best describes the complete sequence of steps occurring during every cycle of PCR?
- The primers hybridize to the target DNA.
- The mixture is heated to a high temperature to denature the double-stranded target DNA.
- Fresh DNA polymerase is added.
- DNA polymerase extends the primers to make a copy of the target DNA.
A) 2, 1, 4 B) 1, 3, 2, 4 C) 3, 4, 1, 2 D) 3, 4, 2 E) 2, 3, 4
A) 2, 1, 4
A researcher needs to clone a sequence of part of a eukaryotic genome in order to express the sequence and to modify the polypeptide product. She would be able to satisfy these requirements by using which of the following vectors?
A) a bacterial plasmid
B) BAC to accommodate the size of the sequence
C) a modified bacteriophage
D) a human chromosome
E) a YAC with appropriate cellular enzymes
E) a YAC with appropriate cellular enzymes
A student wishes to clone a sequence of DNA of ~200 kb. Which vector would be appropriate? A) a plasmid B) a typical bacteriophage C) a BAC D) a plant virus E) a large polypeptide
C) a BAC
Sequencing an entire genome, such as that of C. elegans, a nematode, is most important because
A) it allows researchers to use the sequence to build a “better” nematode, which is resistant to disease.
B) it allows research on a group of organisms we do not usually care much about.
C) the nematode is a good animal model for trying out cures for viral illness.
D) a sequence that is found to have a particular function in the nematode is likely to have a closely related function in vertebrates.
E) a sequence that is found to have no introns in the nematode genome is likely to have acquired the introns from higher organisms.
D) a sequence that is found to have a particular function in the nematode is likely to have a closely related function in vertebrates.
To introduce a particular piece of DNA into an animal cell, such as that of a mouse, you would find more probable success with which of the following methods?
A) the shotgun approach
B) electroporation followed by recombination
C) introducing a plasmid into the cell
D) infecting the mouse cell with a Ti plasmid
E) transcription and translation
B) electroporation followed by recombination
The major advantage of using artificial chromosomes such as YACs and BACs for cloning genes is that
A) plasmids are unable to replicate in cells.
B) only one copy of a plasmid can be present in any given cell, whereas many copies of a YAC or BAC can coexist in a single cell.
C) YACs and BACs can carry much larger DNA fragments than ordinary plasmids can.
D) YACs and BACs can be used to express proteins encoded by inserted genes, but plasmids cannot.
E) All of these are correct.
C) YACs and BACs can carry much larger DNA fragments than ordinary plasmids can.
Which of the following is used to make complementary DNA (cDNA) from RNA? A) restriction enzymes B) gene cloning C) DNA ligase D) gel electrophoresis E) reverse transcriptase
E) reverse transcriptase
Why is it so important to be able to amplify DNA fragments when studying genes?
A) DNA fragments are too small to use individually.
B) A gene may represent only a millionth of the cell’s DNA.
C) Restriction enzymes cut DNA into fragments that are too small.
D) A clone requires multiple copies of each gene per clone.
E) It is important to have multiple copies of DNA in the case of laboratory error.
B) A gene may represent only a millionth of the cell’s DNA.
Pax-6 is a gene that is involved in eye formation in many invertebrates, such as Drosophila. Pax-6 is found as well in vertebrates. A Pax-6 gene from a mouse can be expressed in a fly and the protein (PAX-6) leads to a compound fly eye. This information suggests which of the following?
A) Pax-6 genes are identical in nucleotide sequence.
B) PAX-6 proteins have identical amino acid sequences.
C) Pax-6 is highly conserved and shows shared evolutionary ancestry.
D) PAX-6 proteins are different for formation of different kinds of eyes.
E) PAX-6 from a mouse can function in a fly, but a fly’s Pax-6 gene cannot function in a mouse.
C) Pax-6 is highly conserved and shows shared evolutionary ancestry.
Why are BACs preferred today rather than bacteriophages for making genomic libraries?
A) The BAC carries more DNA.
B) The BAC can carry entire genes and their regulatory elements.
C) Larger BACs are easier to store.
D) The BAC can carry entire genes and their regulatory elements, and larger BACs are easier to store.
E) The BAC carries more DNA, the BAC can carry entire genes and their regulatory elements, and larger BACs are easier to store.
E) The BAC carries more DNA, the BAC can carry entire genes and their regulatory elements, and larger BACs are easier to store.
The reason for using Taq polymerase for PCR is that
A) it is heat stable and can withstand the temperature changes of the cycler.
B) only minute amounts are needed for each cycle of PCR.
C) it binds more readily than other polymerases to primer.
D) it has regions that are complementary to primers.
E) All of these are correct.
A) it is heat stable and can withstand the temperature changes of the cycler.
Why might a laboratory be using dideoxy nucleotides? A) to separate DNA fragments B) to clone the breakpoints of cut DNA C) to produce cDNA from mRNA D) to sequence a DNA fragment E) to visualize DNA expression
D) to sequence a DNA fragment
In order to identify a specific restriction fragment using a probe, what must be done?
A) The fragments must be separated by electrophoresis.
B) The fragments must be treated with heat or chemicals to separate the strands of the double helix.
C) The probe must be hybridized with the fragment.
D) The fragments must be separated by electrophoresis and the fragments must be treated with heat or chemicals to separate the strands of the double helix.
E) The fragments must be separated by electrophoresis, the fragments must be treated with heat or chemicals to separate the strands of the double helix, and the probe must be hybridized with the fragment.
E) The fragments must be separated by electrophoresis, the fragments must be treated with heat or chemicals to separate the strands of the double helix, and the probe must be hybridized with the fragment.
Which of the following modifications is least likely to alter the rate at which a DNA fragment moves through a gel during electrophoresis?
A) altering the nucleotide sequence of the DNA fragment
B) methylating the cytosine bases within the DNA fragment
C) increasing the length of the DNA fragment
D) decreasing the length of the DNA fragment
E) neutralizing the negative charges within the DNA fragment
A) altering the nucleotide sequence of the DNA fragment
DNA fragments from a gel are transferred to a nitrocellulose paper during the procedure called Southern blotting. What is the purpose of transferring the DNA from a gel to a nitrocellulose paper?
A) to attach the DNA fragments to a permanent substrate
B) to separate the two complementary DNA strands
C) to transfer only the DNA that is of interest
D) to prepare the DNA for digestion with restriction enzymes
E) to separate out the PCRs
A) to attach the DNA fragments to a permanent substrate
DNA microarrays have made a huge impact on genomic studies because they
A) can be used to eliminate the function of any gene in the genome.
B) can be used to introduce entire genomes into bacterial cells.
C) allow the expression of many or even all of the genes in the genome to be compared at once.
D) allow physical maps of the genome to be assembled in a very short time.
E) dramatically enhance the efficiency of restriction enzymes.
C) allow the expression of many or even all of the genes in the genome to be compared at once.