18.2

Question 1

Molecular cloning

Answer 1

-GFP naturally found in bioluminescent jellyfish
-Isolated protein can be added to proteins in other spcecies to make them glow

Question 2

Steps of molecular cloning

Answer 2

1. amplify DNA
2. Cut out DNA
3. Isolate cut DNA
4. Insert DNA into new organism

Question 3

Polymerase Chain Reaction (PCR)

Answer 3

-amplify gene of interest
*Denaturation
*Annealing of Primers
*DNA synthesis
*Repeat multiple cycles

Question 4

Denaturation

Answer 4

unwind DNA with heat (no helicase)

Question 5

Annealing of primers

Answer 5

synthetic primers surround DNA of interest

Question 6

DNA synthesis

Answer 6

elongate DNA with DNA polymerase

Question 7

restriction enzymes

Answer 7

-cut amplified DNA at specific locations
* Do not cut into gene of interest
-generate sticky ends

Question 8

gel electrophoresis

Answer 8

-negatively charged DNA runs towards positive charge
-separates DNA fragments by size
*longer fragments travel more slowly
*shorter fragments travel faster
-fluorescent dye allows visualization of fragments

Question 9

ligation

Answer 9

-gene of interest incubated with DNA plasmid
*restriction enzyme cut sites must match
-ligase seals gaps in phosphodiester backbone

Question 10

transformation

Answer 10

-insert GFP plasmid into new bacteria
-Plasmid has:
selection marker
origin of replication

Question 11

genetically modified organisms (GMOs)

Answer 11

ex:transgenic mice that express GFP
ex: glyphosate resistant soy crops