(8) Recombinant DNA Technology Reading Questions

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

Deoxyribonucleoside triphosphates

Which of the following provides the specificity of the PCR reaction? Hint: Most polymerases cannot add nucleotides to DNA without having a short nucleic acid strand in place to which the complementary base can be attached. These short strands, therefore, can be used to provide the specificity of PCR. separated DNA strands primers heating to 94°C Taq polymerase

primers

Which of the following is NOT a vector? Hint: [[Vector Definition]] A vector is a piece of genetic material generated in the laboratory that will transfer specific genes into a cell. viruses transposons plasm ids protoplast fusion

protoplast fusion

A new arrow labeled "lengthens" could be added between ___. Hint: Only enzymes can lengthen nucleic acid strands. "target DNA" -> "primers" "target DNA" -> "DNA strands" "primers" -> "DNA strands" "Taq polymerase" -> "primers"

"Taq polymerase" -> "primers"

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'

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

DNA ligase

A scientist who wants to monitor the gene expression occurring in microbial cells under certain environmental conditions should use which of the following techniques? Hint: [[Techniques of Recombinant DNA Technology]] This procedure utilizes cDNA generated from the RNA of the cell. gene therapy DNA microarrays nucleotide sequencing electroporation

DNA microarrays

Why would a recombinant DNA molecule be inserted into 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 protect the recombinant DNA. Plasmids cannot be isolated outside of a host cell.

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

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

It can withstand the high temperatures associated with PCR.

A plasmid unexpectedly converts to a linear form inside a cell. What will happen to it? Hint: [[The Tools of Recombinant DNA Technology]] Recall that one of the advantages of plasmids is that they are circular DNA molecules. Nothing will happen to it. It will be degraded by cellular enzymes. It will be replicated uncontrollably. It will become inserted into the main chromosome.

It will be degraded by cellular enzymes.

Which of the following methods uses DNA probes? Hint: [[Utility of DNA Probes]] DNA probes can be used to view complementary strands of DNA that have been immobilized on a gel. Southern blot radioactivity electroporation digestion with restriction enzymes

Southern blot

Human proteins can be synthesized in bacteria or yeast through genetic engineering for all of the following reasons EXCEPT: Hint: [[Applications of Recombinant DNA Technology]] Consider that in the past, such proteins had to be isolated from donated blood or from animals. This approach is usually less expensive than traditional methods. The microbially produced protein works better than the original protein. There is no risk of accidentally transferring pathogens. Large quantities of the protein can be made easily.

The microbially produced protein works better than the original protein.

How do restriction enzymes cut DNA sequences? They cut DNA at sequences that have lots of adenine bases. They have the ability to cut DNA randomly. 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 is the function of the primers in PCR? They polymerize free nucleotides to form the new DNA strands. They provide a 3' end for the DNA polymerase. They are the monomer building blocks from which the DNA strand is synthesized. They provide energy for the DNA polymerization reactions.

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

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

If you used a broken thermocycler that could not heat above 75°C, which of the following problems could you expect? Hint: In order for the two strands of DNA to separate, they must be heated up well above 75°C. You would not get any amplification of DNA. You would get more amplification than with a "normal" thermocycler. You would get som~ significant amplification, but less than if you used a "normal" thermocycler. You would get the same amount of amplification as with a "normal" thermocycler.

You would not get any amplification of DNA.

Which of the following would be a useful phenotypic marker on a vector? Hint: [[The Tools of Recombinant DNA Technology]] A phenotypic marker is used to identify cells that have received the gene of interest. antibiotic resistance gene reverse transcriptase ligase restriction enzyme

antibiotic resistance gene

Which of the following is NOT involved in the process of DNA fingerprinting? Hint: [[Applications of Recombinant DNA Technology]] Recall that DNA fingerprinting looks for similarities between patterns of DNA fragments from different cells or individuals. using peR to make multiple copies of a DNA sample gel electrophoresis of DNA fragments addition of fluorescent nucieotides to a DNA sample cutting a DNA sample with restriction enzymes

addition of fluorescent nucieotides to a DNA sample

Put the following events in the construction of a recombinant DNA molecule in the correct order: a. Ligation of desired gene to plasmid DNA. b. Introduction of the plasmid into bacteria c. Restriction enzymes cuts gene of interest and plasmid DNA. d. Growth of cells on an antibiotic-containing medium Hint: [[The Tools of Recombinant DNA Technology]] Think about what tools or materials are needed to begin this process and what the desired result is. c, a, b, d b, c. a, d a, d, c, b d, a, b, c

c, a, b, d

cDNA stands for ___. Hint: [[The Tools of Recombinant DNA Technology]] Recall that cDNA is created by the use of the enzyme reverse transcriptase. copied DNA complementary DNA cloned DNA conventional DNA

complementary DNA

The restriction enzyme EcoRI is used in recombinant DNA technology to ___. alter the outer coverings of Escherichia coli to allow the bacteria to take up recombinant DNA cut DNA at a specific palindromic sequence of N-bases. synthesize DNA copies of mRNA link blunt ends of cleaved double-stranded DNA

cut DNA at a specific palindromic sequence of N-bases.

Vectors are used in recombinant DNA technology to ___. remove an undesirable gene from recipient cell create a DNA microarray form a probe to identify a specific DNA sequence deliver a helpful gene into a recipient cell

deliver a helpful gene into a recipient cell

What is one method by which you could insert DNA from one cell into another cell? Hint: Conjugation does allow for the transfer of genetic material, but it is difficult to control in a laboratory and will not work for eukaryotic cells. conjugation reverse transcriptase restriction enzymes electroporation

electroporation

If a scientist wanted to view the location of a particular species of bacterium within a biofilm, which technique could be used? Hint: [[Applications of Recombinant DNA Technology]] To locate that species, one would have to find something unique about the species and then find a way to make that trait obvious. electroporation fluorescent in situ hybridization (FISH) nucleotide sequencing DNA fingerprinting

fluorescent in situ hybridization (FISH)

Electroporation is a technique used for ___. Hint: [[Techniques of Recombinant DNA Technology]] Consider that electroporation involves an electrical current that creates holes in the cell membrane. inserting DNA into cells separating DNA molecules by size determining the base sequence of a DNA molecule generating fragments of DNA

inserting DNA into cells

The small size of a genetic vector is an advantage because ___. Hint: [[The Tools of Recombinant DNA Technology]] Recall that a genetic vector is a mechanism for inserting a useful gene into a host cell. they can be degraded more quickly than larger DNA molecules they are lighter than larger DNA molecules larger molecules would be too fragile to manipulate in the laboratory they are small enough to enter the cell undetected

larger molecules would be too fragile to manipulate in the laboratory

A Southern blot is used to ___. Hint: [[Techniques of Recombinant DNA Technology]] Recall that Southern blots involve the transfer of DNA fragments from an agarose gel to a more stable nitrocellulose membrane. locate DNA sequences of interest in a collection of DNA fragments separate a group of DNA fragments by size determine the base sequence of a DNA molecule create millions of copies of a target DNA sequence

locate DNA sequences of interest in a collection of DNA fragments

Bt toxin is important in which of the following agricultural applications? Hint: [[Applications of Recombinant DNA Technology]] Bt toxin exerts its activity only after it has entered the intestinal tract of an insect. salt tolerance freeze resistance herbicide tolerance pest resistance

pest resistance

Which of the following enzymes is used to make specific cuts in DNA molecules? Hint: [[The Tools of Recombinant DNA Technology]] These enzymes are bacterial in origin and they make one of two major types of cuts in DNA molecules. reverse transcriptase probes DNA ligase restriction enzymes

restriction enzymes

Gene replacement therapy uses polymerase chain reaction (PCR). PCR is used in recombinant DNA technology to ___. synthesize millions of copies of a DNA molecule synthesize RNA copies of messenger RNA separate DNA fragments in agarose gel insert recombinant DNA molecules into a recipient cell

synthesize millions of copies of a DNA molecule

All of the following pieces of information are conveyed by the name of a restriction enzyme EXCEPT ___. Hint: [[The Tools of Recombinant DNA Technology]] A common restriction enzyme is EcoRI. strain of the bacterium in which it was discovered. the order of its discovery compared to other restriction enzymes from that bacterium. the species of the bacterium in which it was discovered. the type of cut it makes.

the type of cut it makes.

Four different fluorescent dyes are used in automated DNA sequencing because ___. Hint: [[Applications of Recombinant DNA Technology]] Consider that an automated DNA sequencer will actually be determining the sequence of the four different dyes. DNA molecules are cut into four pieces during the process there are four different types of DNA that can be sequenced using the procedure there are four different bases in DNA the process involves four different steps

there are four different bases in DNA

Protoplasts are valuable in techniques that are used to insert DNA in cells because ___. Hint: [[Techniques of Recombinant DNA Technology]] Recall that cells of fungi and algae must be converted to protoplasts before they can be used in some of these techniques. they contain no DNA of their own. they cannot degrade the DNA once it has entered the cell. they reproduce more quickly than other types of cells. they have no cell wall to interfere with the entry of DNA into the cell.

they have no cell wall to interfere with the entry of DNA into the cell.