Lecture 6 #2

Front 1

Restriction- fragment- length- polymorphisms (RFLPs):

Back 1

1) are restriction enzyme-generated DNA fragments of different lengths

2) are cut from the same region of the genome

3) are re separated using agarose gel electrophoresis

4) can be analyzed using a hybridization-based method called Southern-blot analysis

Front 2

Genetic engineering of bacteria has been use to produce which protein?

Back 2

a. human growth hormone to treat human growth disorders

b. human insulin to treat diabetes

c. tissue plasminogen activator to dissolve blood clots that cause heart attacks

d. a vaccine against foot-and-mouth disease of cattle

Front 3

In the figure above, which lane shows a very large, linear DNA fragment that was cut by two enzymes?

Back 3

5

Front 4

Which enzyme did I use if I cut a DNA fragment and ended up with a double stranded sequence that looks like this: AATTC---------G G---------CTTAA

Back 4

Eco RI

Front 5

Identify the correct steps and the correct order for cloning a gene of interest.

Back 5

1) Use restriction endonucleases to cut the gene of interest and the target DNA.

2)Incubate the combined DNA fragments with the liagase enzyme.

3)Mix the postrestriction endonuclease gene of interest with the postrestriction endonuclease cloning plasmid.

4) Transform the bacteria.

5) Spread bacteria on medium containing X-gal and ampicillin.

Front 6

What is used is used as a vector for making transgenic plants, in much the same way as cloning vectors introduce genes into bacteria?

Back 6

the Ti plasmid

Front 7

In restriction enzymes:

Back 7

1) Restriction enzymes recognize specific restriction sites

2) Restriction enzymes used in cloning produce DNA fragments with single-stranded ends.

3) DNA fragments produced by a restriction enzyme are known as restriction fragments.

4) Bacterial enzymes can be used to generate DNA fragments that can be joined to produce recombinant DNA molecules.

Front 8

A set of clones that collectively contains a copy of every DNA sequence in a genome is called a ________.

Back 8

genomic DNA library

Front 9

Cloning plasmids generally contain genes for which two traits?

Back 9

beta-galactosidase enzyme production and ampicillin resistance

Front 10

If a restriction endonuclease cuts a linear DNA three times, how many fragments should there be after the DNA is subjected to electrophoresis through an agarose gel?

Back 10

4

Front 11

In ________, a gene of interest is identified in the set of clones when it base-pairs with a short, single-stranded complementary DNA or RNA molecule.

Back 11

DNA hybridization

Front 12

In the figure above, which DNA fragment is the smallest?

Back 12

E

Front 13

If multiple fragments from a genome have been cloned into bacteria, how can a researcher identify the bacteria that contains the particular sequence they are interested in?

Back 13

The DNA purified from individual colonies of bacteria is probed with a short, single stranded DNA fragment that is complementary to the gene of interest.

Front 14

Recombinant DNA is:

Back 14

DNA fragments from two or more sources that have been joined together.

Front 15

What is/are true of the polymerase chain reaction?

Back 15

1) PCR allows for the mass production of a specific DNA sequence without cloning.

2) PCR allows scientists to study DNA from samples of finite amounts, such as from fossils, or small tissue samples left at a crime scene.

3) PCR cannot be used to clone DNA of unknown sequence

4) PCR permits scientists to target a single copy of a single gene among millions of other sequences.

Front 16

Why is it important to have an antibiotic resistance gene in the cloning plasmid?

Back 16

It provides a way for researchers to sort the bacteria that have the cloning plasmid from the bacteria that don't have the cloning plasmid.