Chapter 18 Recombinant Dna And Biotechnology

Front 1

What is the specific name used for bacterial enzymes that are used to cut DNA?

Back 1

Restriction endonucleases (RE)

Front 2

Restriction enzymes recognize ______ DNA sequences. What exactly are these?

Back 2

palindromic
A sequence of DNA that is read the same way backwards and forwards. For example:
GAATTC
CTTAAG

Front 3

What are the different types of "cuts" a restriction enzyme can make?

Back 3

1. Straight cuts
2. Staggered cuts

Front 4

What is a "sticky end"?

Back 4

When restriction enzymes make a staggered cut, each fragement carries single stranded "over hang" at the location of the cut.

Front 5

How do sticky ends combine?

Back 5

The base pairs of one sticky end combine with the complementary base pairs of another sticky end

Front 6

It is not possible for fragmented DNA from DIFFERENT organisms to be joined. T or F?

Back 6

False

Front 7

What catalyzes the formation of covalent bonds between adjacent nucleotides at the ends of the fragements?

Back 7

DNA ligase
This is basically the "glue" that joins them together to form a single larger molecule

Front 8

A DNA molecule that is derived from at least two genetic sources in a labratory is called what?

Back 8

Recombinant DNA

Front 9

One of the main purposes of recombinant DNA is to make ______ of genes?

Back 9

clones

Front 10

The cloning of recombinant DNA by inserting it into a host cell OTHER THAN an animal cell is called what?

Back 10

Transformation

Front 11

The cloning of recombinant DNA by inserting it into an ANIMAL host cell is called what?

Back 11

Transfection

Front 12

When cloning cells, almost every "host" cell that is injected becomes transformed. T or F?

Back 12

False
Usually only a few are transformed

Front 13

A host cell or organism that contains recombinant DNA is referred to as what?

Back 13

Transgenic

Front 14

How does a biologist determine which hosts cells contain the newly planted sequence?

Back 14

Through selectable markers.
For example, in our lab, the gfp glowing protien was a selectable marker. It allowed us to easily IDENTIFY that the bacteria we were looking at contained that specific gene

Front 15

How exactly do biologists insert recombinant DNA into a host cell? Give 4 examples.

Back 15

1. Chemically treat the host: making the membrane more permeable so that DNA can diffuse into it
2. Electroporation: short electric shock which creates temporary pores in the membrane for DNA to enter
3. Viruses can carry it into cells
4. Transformed plants
5. Transgenic animals: injecting the DNA into the nuclei of fertilized eggs

Front 16

What happens after DNA is succesfully inserted into a host cell?

Back 16

Host cell divides and new DNA replicates. Clones!

Front 17

The challenge of inserting DNA is getting it to replicate. For this to happen, the inserted DNA must become part of a segment of DNA with an ORI. What is this called?

Back 17

A replicon

Front 18

An newly inserted DNA can become part of a replicon in one of two ways. What are they?

Back 18

1. It may be inserted into a host chromosome near its ORI
2. It can enter the host cell as part of a carrier DNA sequence called a vector

Front 19

What makes are really good vector and why?

Back 19

Plasmid
1. Small and easy to manipulate
2. Have one or more restriction enzyme recognition sequences
3. Have a bacterial ORI

Front 20

What is the drawback of plasmids.

Back 20

It can only hold up to 10,000 base pairs. This is fairly small, so therefore cannot hold the large eukaryotic genes with very large base sequences.

Front 21

A virus can be a vector. T or F?

Back 21

True: these viruses are called bacteriophage

Front 22

What is a reporter gene?

Back 22

A selectable marker that has an EASILY observable gene expression. For example: the gfp glowing gene from our lab!

Front 23

Explain what is happening in this picture.

Back 23

1. This is a plasmid and it is acting as a vector
2. The GFP gene has been spliced into it
3. This plasmid is then inserted into host bacteria
4. The bacteria replicates (clones)
5. Was it succesful in replicating??
6. Yes! because we can see that each one of the clones who has the spliced-in gene glows under UV light

Front 24

A collection of DNA fragments that together comprise the genome of an organism is called what?

Back 24

A genomic library

Front 25

This illustration is describing how a genomic library is constructed. What is happening in this diagram?

Back 25

1. Small fragments of the genome DNA is cut up
2. These fragments are inserted into vectors (plasmids)
3. Plasmids are inserted into a host cell
4. This cell produces a COLONY of recombinant cells

Front 26

A smaller DNA library (as opposed to a genomic library) include only the genes that transcribe a particular tissue. What type of DNA aids in this process?

Back 26

cDNA (complementary DNA)

Front 27

This picture describes how a library is made using cDNA. What is happening in this diagram?

Back 27

1. A strand of mRNA is isolated from a cell
2. Through complementary base pairing, cDNA copies are made (reverse transcriptase)
3. cDNA is spliced into a vector (plasmid)
4. Vector injected into host cell
5. Host cell replicates and cDNA library is created

Front 28

The use of living cells or organisms to produce materials useful to people is called what?

Back 28

Biotechnology

Front 29

Give several examples of biotechnology.

Back 29

1. Yeasts to brew beer and wine
2. Bacteria to make cheese and yogurt
3. Microbes to produce antibiotics, alcohol, etc

Front 30

The production of pharmaceuticals in farm animal or plants is called what?

Back 30

Pharming

Front 31

Give an example of pharming (hint: cows).

Back 31

1. A gene that encodes a USEFUL protien can be inserted next to the promoter of the gene in cows that encodes for the protein lactoglobulin (found in milk)
2. Any cow that carries this recombinant DNA will produce large amounts of the USEFUL protein into their milk.
3. This USEFUL protien can then be separated from the milk and used for medical purposes.

Front 32

The use of organisms to remove contaminants from the environment is called what?

Back 32

Bioremediation

Front 33

Give an example of bioremediation.

Back 33

Composting: Involves the use of bacteria to break down large molecules and proteins in waste products such as wood chips, paper, straw and kitchen scraps.

Front 34

Transgenic organisms can be used to clean up environmental contaminants such as oil spills. T or F?

Back 34

True