Chapter 8 Recombinant DNA Technology

Which of the following attaches the target gene to a desired location? Plasmids DNA ligase Restriction enzymes Chromosomal DNA

DNA ligase

Which of the following would be most useful in following gene expression in a yeast cell? Southern blot PCR DNA microarray restriction enzymes

DNA microarray

In which direction does DNA polymerase synthesize the new DNA strand? Both 5' to 3' and 3' to 5' 5' to 3' 3' to 5'

5' to 3'

Synthetic nucleic acids are useful as primers for PCR. DNA probes. antisense RNAs. DNA probes and antisense RNAs. DNA probes, primers, and antisense RNAs.

DNA probes, primers, and antisense RNAs.

What provides the energy for DNA polymerization in a PCR reaction? Primers DNA polymerase Deoxyribonucleoside triphosphates Template DNA

Deoxyribonucleoside triphosphates

Which type of molecule serves as a template during transcription, and which type of molecule is synthesized? DNA is the template during transcription, and RNA is synthesized. RNA is the template during transcription, and DNA is synthesized. DNA is the template during transcription, and proteins are synthesized. RNA is the template during transcription, and proteins are synthesized.

DNA is the template during transcription, and RNA is synthesized. During transcription, a DNA sequence is used as a template to synthesize an RNA strand complementary to the DNA. The RNA strand is then used as a template to synthesize protein.

Chemical and physical agents can cause changes in nucleotide squences in DNA. Such agents are polymerase chain reactions. restriction enzymes. reverse transcriptase. mutagens.

Mutagens, Mutagens are chemical or physical agents that can change DNA sequences.

What characteristic(s) of gel electrophoresis make it an effective technique in genetic research? Positively charged DNA travels toward the negative electrode. Negatively charged DNA travels toward the positive electrode. Large DNA is attracted to the positive end of the gel Small DNA is attracted to the negative end, while larger DNA is not.

Negatively charged DNA travels toward the positive electrode. DNA is negatively charged and is attracted to the positive end of the gel.

Four different fluorescent dyes are used in automated DNA sequencing because __________. the process involves four different steps DNA molecules are cut into four pieces during the process there are four different nucleotides in DNA there are four different types of DNA that can be sequenced using the procedure

there are four different nucleotides in DNA

In the laboratory, recombinant DNA technologists use restriction enzymes __________. as genes to cut specific genes from one organism for the purpose of inserting the genes into the DNA of another organism only as a last resort to destroy pathogenic bacteria within a human cell

to cut specific genes from one organism for the purpose of inserting the genes into the DNA of another organism. Yes. Restriction enzymes make it possible to excise a specific gene from a sequence of DNA, and another enzyme, ligase, glues it to a new piece of DNA.

steps of PCR on the following figure. Indicate the temperature of the reaction at each numbered step.

1. 1.Step 1, denaturation: 94°C; step 2, priming: DNA primer, deoxyribonucleotide triphosphates, DNA polymerase, cool to 65°C; step 3, extension: same reagents as step 2, 72°C; step 4, repeat.

What is the temperature used for the extension step? 72 °C 60 °C 94 °C

72 °C

What is the sequence of the temperatures of a typical PCR reaction? 60 °C, 72 °C, 94 °C 94 °C, 60 °C, 72 °C 72 °C, 94 °C, 60 °C 72 °C, 60 °C, 94 °C 94 °C, 72 °C, 60 °C

94 °C, 60 °C, 72 °C`

Where could a research scientist find copies of all the genes of a microbe without having to isolate them from the microbe? CRISPR-Cas cDNA Restriction enzymes Gene library

A gene library is a collection of clones containing fragments (typically single genes) of a genome.

Which of the following would not be a potential medical application of recombinant DNA technology: gene library, genetic mapping, gene therapy, xenotransplantaton? Gene therapy Genetic mapping Gene library Xenotransplantation

A gene library is a collection of clones containing fragments (typically single genes) of a genome; it isn't necessarily part of medicine.

What is the difference between a missense mutation and a nonsense mutation? A missense mutation codes for an amino acid that is incorrect for the polypeptide, while a nonsense mutation changes a codon for an amino acid into a stop codon. A missense mutation changes the code for an amino acid into a stop codon, whereas a nonsense mutation changes the code for one amino acid into the code for a different one. A missense mutation is a change in nucleotide sequence that does not change the amino acid sequence in the polypeptide, while a nonsense mutation changes the codon to code for a different amino acid. A missense mutation codes for an amino acid that is incorrect for the polypeptide, whereas a nonsense mutation changes a stop codon into a codon for an amino acid.

A missense mutation codes for an amino acid that is incorrect for the polypeptide, while a nonsense mutation changes a codon for an amino acid into a stop codon.

What is the difference between a missense mutation and a nonsense mutation? A missense mutation codes for an amino acid that is incorrect for the polypeptide, whereas a nonsense mutation changes a stop codon into a codon for an amino acid. A missense mutation codes for an amino acid that is incorrect for the polypeptide, while a nonsense mutation changes a codon for an amino acid into a stop codon. A missense mutation is a change in nucleotide sequence that does not change the amino acid sequence in the polypeptide, while a nonsense mutation changes the codon to code for a different amino acid. A missense mutation changes the code for an amino acid into a stop codon, whereas a nonsense mutation changes the code for one amino acid into the code for a different one.

A missense mutation codes for an amino acid that is incorrect for the polypeptide, while a nonsense mutation changes a codon for an amino acid into a stop codon. A missense mutation results in a change from one amino acid to a different one, whereas a nonsense mutation results in the change of an amino acid codon into a stop codon. Nonsense mutations almost always result in nonfunctional proteins because they introduce premature stop codons. Missense mutations may result in functional proteins, depending on which amino acid is affected.

A scientist wants to add DNA to a particular genome using a vector. One vector she might use is: injection a mutagen. a plasmid. reverse transcription.

A plasmid, Plasmids are circular, extrachromosomal, DNA molecules that are small, stable, and can contain promoters

Which of the following devices is used for PCR? a thermocycler an electrophoresis chamber a nucleic acid creation machine a DNA sequencer a gene gun

A thermocycler

What is the name of genetic tools that can deliver genes into cells? Virus genomes Vectors Transposons Plasmids

A vector is any nucleic acid molecule that can deliver a gene into a cell.

Baking bread, creating a genetically modified vegetable, using a microbe to make human insulin . . . Which of these is NOT considered "biotechnology"? Using a microbe to make human insulin Creating a GMO vegetable Baking bread All of these are biotechnology

All of these examples are biotechnology. Biotechnology simply means using microbes to make a practical product.

A scientist wants to insert a gene into several different organisms at several different sites. The best type of restriction enzymes would be ones that produce: vectors. blunt ends. sticky ends. DNA probes.

Blunt ends.(Blunt-ended DNA fragments can be joined even if they are produced by different restriction enzymes.)

Which of the following statements concerning sticky-ended fragments is most likely to be true? The sticky-ended fragments are much harder to join together than blunt-ended fragments. Complementary sticky-ended fragments were probably cut by the same restriction enzyme. Sticky-ended fragments have the ability to join any other sticky fragment, regardless of whether or not it is complementary. Sticky-ended fragments will never be more than two nucleotide bases long.

Complementary sticky-ended fragments were probably cut by the same restriction enzyme. Yes. Sticky-ended DNA fragments cut by the same restriction enzyme will always be complementary to one another. Sticky-ended fragments cut by two different restriction enzymes will most likely be different and thus not complementary to one another.

Which type of molecule serves as a template during transcription, and which type of molecule is synthesized? RNA is the template during transcription, and DNA is synthesized. DNA is the template during transcription, and proteins are synthesized. DNA is the template during transcription, and RNA is synthesized. RNA is the template during transcription, and proteins are synthesized.

DNA is the template during transcription, and RNA is synthesized. During transcription, the DNA sequence of a gene is used as a template to synthesize an RNA strand complementary to the DNA.

Which technique would be most useful in diagnosing an infection? Southern blotting DNA microarray DNA sequencing Heat shocking

DNA microarrays made with sequences from several pathogens can reveal the presence of a pathogen causing an infection.

How are restriction enzymes named? Restriction enzymes are usually titled according to the nucleotide sequence of the restriction site upon which they act. Restriction enzymes are usually named after the scientist that discovers them. The naming of a restriction enzyme involves several letters corresponding to an amino acid and a Greek letter implying the type of organism on which they act. Each is named with letters referring to the genus, species, and strain of the source prokaryote and a Roman numeral denoting the order of its discovery.

Each is named with letters referring to the genus, species, and strain of the source prokaryote and a Roman numeral denoting the order of its discovery

Which technique would be useful for locating a particular gene in living cells? Next-generation sequencing PCR DNA microarrays FISH

FISH

Which technique would be useful for locating a particular gene in living cells? Next-generation sequencing PCR FISH DNA microarrays

FISH or fluorescent in situ hybridization, is used to label living cells with a fluorescent marker.

Restriction enzymes are useful only on synthetic DNA molecules. True/False

False

T/F A thermocycler separates molecules based on their size, shape, and electrical charge.

False, Electrophoresis separates molecules based on their size, shape, and electrical charge.

T/F Gel electrophoresis is used in DNA microarrays.

False, Gel electrophoresis is used in Southern blotting

T/F Restriction enzymes inhibit the movement of DNA at specific sites.

False, Restriction enzymes cut DNA at specific sites.

Which of the following techniques is used regularly in the study of genomics? Clones are selected using a vector with two genetic markers. Genes are inserted to produce an antigenic protein from a pathogen. Fluorescent nucleotide bases are sequenced. Defective organs are replaced with those made in animal hosts.

Fluorescent nucleotide bases are sequenced.

Why would a recombinant DNA molecule be inserted into a host cell? It can protect the recombinant DNA. Plasmids cannot be isolated outside of a host cell. It can be copied, transcribed, and translated into a desired protein. Restriction enzymes can only be used inside of a cell.

It can be copied, transcribed, and translated into a desired protein.

Why is DNA polymerase from Thermus aquaticus ideal for PCR? It does not require energy to polymerize DNA. It can withstand the high temperatures associated with PCR. It can synthesize DNA 5' to 3' and 3' to 5'. It does not require primers.

It can withstand the high temperatures associated with PCR.

Which of the following statements is true concerning recombinant DNA technology? It will replace biotechnology in the future. It is a single technique for genetic manipulation. It is useful in manipulating genotypes but not phenotypes. It involves modification of an organism's genome.

It involves modification of an organism's genome.

What is the purpose of a primer during DNA replication? It catalyzes the addition of nucleotides to the growing DNA strand. It provides an initial 3ʹ hydroxyl group for DNA polymerase to add nucleotides. It attaches newly synthesized DNA strands to each other. It breaks the hydrogen bonds between the two DNA strands, separating them.

It provides an initial 3ʹ hydroxyl group for DNA polymerase to add nucleotides. A primer is a short stretch of nucleotides that is complementary to the DNA template strand and binds to it. A primer provides a 3ʹ hydroxyl group at the end of the nucleotide to which DNA polymerase can add additional nucleotides complementary to the template strand.

What is the purpose of a primer during DNA replication? It catalyzes the addition of nucleotides to the growing DNA strand. It attaches newly synthesized DNA strands to each other. It provides an initial 3′ hydroxyl group for DNA polymerase to add nucleotides. It breaks the hydrogen bonds between the two DNA strands, separating them.

It provides an initial 3′ hydroxyl group for DNA polymerase to add nucleotides. DNA polymerase can only add nucleotides to an existing 3′ hydroxyl group.

Which recombinant DNA technique is used to replicate copies of a DNA molecule? PCR gel electrophoresis electroporation reverse transcription

PCR

What are plasmids? Plasmids are small, usually circular, molecules of DNA often found in prokaryotes that replicate independently. Plasmids are small, usually linear molecules of DNA that are critical for cellular survival. Plasmids are large, usually linear molecules of DNA, found mainly in eukaryotes. Plasmids are large, usually circular molecules of DNA that are critical for cellular survival.

Plasmids are small, usually circular, molecules of DNA often found in prokaryotes that replicate independently Plasmids are small, usually circular molecules of DNA. They are often, but not exclusively, found in prokaryotes. They replicate independently from the genome of the cell. They are not required for cellular survival, though naturally occurring plasmids can code for beneficial characteristics, such as for resistance to antibiotics or for toxins. In this chapter you will learn how plasmids can be modified for research and biotechnology purposes.

What are plasmids? Plasmids are large, usually linear molecules of DNA, found mainly in eukaryotes. Plasmids are large, usually circular molecules of DNA that are critical for cellular survival. Plasmids are small, usually circular, molecules of DNA often found in prokaryotes that replicate independently. Plasmids are small, usually linear molecules of DNA that are critical for cellular survival.

Plasmids are small, usually circular, molecules of DNA often found in prokaryotes that replicate independently. Plasmids are independently replicating molecules of DNA often found in prokaryotes in addition to the genome.

What process is used to "amplify" DNA, making large numbers of copies from a small sample? Thermocycler Gene libraries Priming Polymerase chain reaction

Polymerase chain reaction

Which technique or tool of recombinant DNA technology makes many copies of a DNA molecule? Electrophoresis Restriction enzymes Reverse transcriptase (RT) Polymerase chain reaction (PCR)

Polymerase chain reaction (PCR) PCR is a method whereby numerous copies of a DNA molecule is made.

Small single strand DNA molecules with a fluorescent molecule attached that are used to identify a clone containing a specific gene fragment is called a_____ promoter. primer. peptide. probe. plasmid.

Probe

Mutagens are useful in biotechnology research for removing undesirable traits from microbes. producing DNA fragments for cloning. selecting genetic mutants resistant to radioactivity. producing organisms with altered phenotypes. producing new organisms which have beneficial traits from two or more organisms.

Producing organisms with altered phenotypes.

Which of the following enzymes can recognize and make cuts in specific DNA sequences? DNA ligase reverse transcriptase DNA polymerase restriction enzymes

Restriction enzymes

What function do restriction enzymes perform? They read mRNA and write cDNA. They replace RNA with DNA on a lagging strand. They cut DNA at specific sequences. They splice DNA into a lagging strand.

Restriction enzymes cut DNA at "restriction sites" that are unique to each restriction enzyme.

Which viral enzyme is used to copy nucleotide sequences to make cDNA? DNA polymerase I Helicase Reverse transcriptase DNA polymerase III

Reverse transcriptase is a viral enzyme that copies mRNA into DNA that lacks introns (cDNA).

How do the strands separate during PCR? The DNA polymerase breaks the hydrogen bonds between the two strands. The primers separate the strands during the annealing step The cycling of the temperatures breaks the hydrogen bonds between the two strands. The high heat of the denaturation step breaks the hydrogen bonds between the two strands.

The high heat of the denaturation step breaks the hydrogen bonds between the two strands.

What is a thermocycler? The machine that controls the heat of the reaction, cycling between the different temperatures of the different steps during PCR The name for the DNA primers used in a PCR reaction The process of cycling through the different temperatures of a PCR reaction 30 times The special DNA polymerase, used in a PCR reaction, that can tolerate the high temperatures

The machine that controls the heat of the reaction, cycling between the different temperatures of the different steps during PCR

All restriction sites are palindromic. What does this mean? It means that the restriction sites are the same for both prokaryotes and eukaryotes. It means that a restriction site may be targeted by any of the restriction enzymes. The restriction sites are always five nucleotide base pairs in length. The nucleotide sequences at the restriction sites are the same when read forward or backward.

The nucleotide sequences at the restriction sites are the same when read forward or backward. Yes. It means that the nucleotide sequences of two complementary strands are exactly repeated.

All of the following pieces of information are conveyed by the name of a restriction enzyme EXCEPT __________. the species of the bacterium in which it was discovered the genus of bacterium in which it was discovered the type of cut it makes the order of its discovery compared to other restriction enzymes from that bacterium

The type of cut it makes

Although blunt-ended DNA fragments are more difficult to join back together than are sticky ends, blunt ends have an advantage in that __________. Once joined to another blunt-ended fragment, the DNA can never be cut again They form many more hydrogen bonds than sticky-ended fragments Blunt-ended fragments can be used to make recombinant DNA between two different species They can be recombined with blunt fragments produced by different restriction enzymes

They can be recombined with blunt fragments produced by different restriction enzymes. Yes. Blunt-ended fragments are non-specific and can be joined to any other blunt-ended fragment, regardless of the restriction enzymes used.

How do restriction enzymes cut DNA sequences? They have the ability to cut DNA randomly. They cut DNA at sequences that have lots of adenine bases. They cut DNA at sites, called recognition sites, that have specific nucleotide sequences.

They cut DNA at sites, called recognition sites, that have specific nucleotide sequences.

What action do restriction enzymes employ? They glue two pieces of DNA back together. They cut DNA. They inhibit DNA from being transcribed. They facilitate the production of hydrogen bonds.

They cut DNA. Yes. Restriction enzymes work by cutting DNA at sites with very specific nucleotide sequences.

What role do restriction enzymes serve in the prokaryotes that produce them? They provide a means by which different prokaryotic species can share genes. They destroy viral DNA. They are involved in bacterial apoptosis or cell suicide. They play a role in DNA synthesis during reproduction.

They destroy viral DNA Yes. The restriction enzymes cause the destruction of viral DNA that might otherwise harm the bacterium or archaeon.

What is the function of the primers in PCR? They provide energy for the DNA polymerization reactions. They are the monomer building blocks from which the DNA strand is synthesized. They provide a 3' end for the DNA polymerase. They polymerize free nucleotides to form the new DNA strands.

They provide a 3' end for the DNA polymerase.

In general, how might recombinant DNA technology be used to prevent a genetic disorder caused by a mutation in a single gene?

To insert a desirable gene, remove an undesirable gene, or replace a defective gene with a functioning gene

Reverse transcriptase synthesizes a DNA molecule from an RNA template. True/False

True

Southern blotting is a technique that can be used to identify microbes that cannot be cultured. True/False

True

T/F Protoplast fusion is often used in the genetic modification of plants.

True

T/F Restriction enzymes act at specific nucleotide sequences within a double-stranded DNA molecule.

True

Which of the following is false concerning vectors in recombinant DNA technology? Vectors are small enough to manipulate outside a cell. Vectors contain a recognizable genetic marker. Vectors survive inside cells. Vectors must contain genes for self-replication.

Vectors must contain genes for self-replication.

The restriction enzyme KpnII probably originated __________ as the second discovered restriction enzymes from Klebsiella pneumoniae bacterium from Escherichia coli, one of the most widely studied bacteria from a virus as the product of recombination between a bacterium and a plant

as the second discovered restriction enzymes from Klebsiella pneumoniae bacterium.Yes. KpnI is the name for the first restriction enzyme discovered from K. pneumoniae; KpnII was discovered second.

A DNA gene synthesized from an RNA template is . reverse transcriptase complementary DNA recombinant DNA probe DNA

complementary DNA

The CRISPR-associated enzyme Cas9 . cuts RNA at a specific nucleotide sequence cuts DNA at a specific nucleotide sequence is likely derived from a bacteriophage All of the above are correct.

cuts DNA at a specific nucleotide sequence

Which application of recombinant DNA technology involves replacing a nonfunctional, mutated gene with a functional gene? gene therapy functional genomics genetic screening protein synthesis

gene therapy

Which of the following procedures would be used to introduce DNA into a single mouse cell? microinjection gene gun Southern blotting electroporation protoplast fusion

microinjection

After scientists exposed cultures of Penicillium to agents X, Y, and Z, they examined the type and amount of penicillin produced by the altered fungi to find the one that is most effective. Agents X, Y, and Z were probably . recombinant cells competent mutagens phages

mutagens

Bt toxin is important in which of the following agricultural applications? pest resistance herbicide tolerance resistance to freezing salt tolerance

pest resistance

A DNA microarray consists of . a series of clones containing the entire genome of a microbe recombinant microbial cells restriction enzyme fragments of DNA molecules single-stranded DNA localized on a substrate

single-stranded DNA localized on a substrate