CH 9 MASTERING - BIOTECHNOLOGY & DNA TECHNOLOGY

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Which of the following places the steps in the PCR procedure in the correct order? 1) Incubate at 94C to denature DNA strands 2) Incubate at 72C for DNA synthesis 3) Incubate at 60C for primer hybridization

1,3,2

Which of the following best describes how recombinant DNA technology currently helps patients who do NOT produce adequate amounts of growth hormone (hGH) - a condition that otherwise leads to stunted growth?

Bacteria now produce hGH

In the figure, the bacteria transformed with the recombinant plasmid and plated on media containing ampicillin and X-gal will

form white, ampicillin-resistant colonies.

Self-replicating DNA used to transmit a gene from one organism to another is a

vector

Arrange the following steps from a typical genetic modification procedure into their correct sequence.

1. Isolate bacterial plasmid 2. Select gene of interest + insert into plasmid 3. Plasmid taken up by bacterium 4. Bacteria replicate 5. Protein product harvested from bacteria AND cloned gene harvested from bacteria

In the figure, after digestion with the appropriate restriction enzyme, what is the smallest piece containing the entire ampicillin-resistance (amp) gene?

1.50 kbp

How many pieces with EcoRI produce from the plasmid shown in the figure?

2

What is the temperature used for the extension step?

72C

What is the sequence of the temperatures of a typical PCR reaction?

94C, 60C, 72C

Which of the following best describes a clone in the context of genetic modification procedures?

A culture of genetically identical cells

Which of the following best describes why a vector is used in genetic modification procedures?

Cells usually won't copy an isolated gene sequence

Which of the following attaches the target gene to a desired location?

DNA ligase

Foreign DNA can be inserted into cells using a variety of different methods. Which method involves the formation of microscopic pores in the cell's membrane?

Electroporation

Which enzyme would cut this strand of DNA? GCATGGATCCCAATGC

Enzyme Recognition BamHI G GATCC CCCTAG ^ g

Nearly all cells, including E. coli and yeast, naturally take up DNA from their surroundings without chemical treatment.

False

Why would a recombinant DNA molecule be inserted into a host cell?

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

You want to determine whether a person has a certain mutant gene. The process involves using a primer and a heat-stable DNA polymerase. This process is

PCR

Recombinant DNA techniques typically involve generating a clone. Why?

Producing a clone generates many copies of the gene of interest

Which of the following is an application that uses PCR?

Sequencing a gene, diagnosing a disease, and providing enough DNA for cloning into another organism

How do the strands separate during PCR?

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

A source of heat-stable DNA polymerase is

Thermus aquaticus

What is a thermocycler?

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

Which statement best describes restriction enzymes?

They are important for cloning applications because they can be used to cut out DNA at specific nucleotide sequences.

What is the end goal of PCR?

To quickly increase the number of copies of a specific DNA sequence

Which of the following methods would be used to introduce the plasmid shown in the figure into E.coli?

Transformation

Which of the following processes is not involved in making cDNA?

Translation

One of the first commercial successes of recombinant DNA technology was the production of human insulin using genetically engineered E.coli.

True

Why is baker's yeast useful for expressing genetically engineered genes?

Yeast cells are eukaryotic and so would likely be successful in expressing eukaryotic genes.

A restriction fragment is

a segment of DNA

In the figure, the marker genes used for selecting recombinant DNA are

ampR and lacZ

Restriction enzymes function as

bacterial enzymes that destroy phage DNA

A population of cells carrying a desired plasmid is called a

clone

An advantage of using Saccharomyces cerevisiae over Escherichia coli to produce a human protein is that

it will be more likely to continuously secrete the protein.

An advantage of cDNA over genomic DNA is that it

lacks introns

While ______ is/are responsible for the diversity of life, ________ is/are responsible for shaping a population with organisms possessing characteristics that enhance survival.

mutations; selections

The use of "suicide" genes in genetically modified organisms is designed to

prevent the growth of the modified organisms in the environment.

Assume you have discovered a cell that produces a lipase that works in cold water for a laundry additive. You can increase the efficiency of this enzyme by changing one amino acid. This is called

site-directed mutagenesis

PCR can be used to identify an unknown bacterium because

the DNA primer is specific to particular target DNA sequences.


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