Recombinant DNA Technology Flashcards
1
Q
Enzymes used in recombinant DNA technology are used as tools to manipulate DNA to:
A
- make copies of DNA
- cut DNA molecules into shorter fragments
- join DNA molecules together
In combinations that do not exist in nature
2
Q
What are Restriction Endonucleases
A
Enzymes that bind to a DNA molecule at a particular recognition sequence and make specific double stranded cut at or near that sequence
3
Q
What do types I & III restriction endonucleases do
A
They cut randomly around target sequence.
Limited use as unpredictable in the position it cuts at.
4
Q
What does type II restriction endonuclease do
A
Always cuts at the same position within or near recognition site.
Predictable activity.
5
Q
Characteristics of blunt ends
A
- Double stranded at either ends
- Antiparallel to eachother
6
Q
Characteristics of sticky ends
A
- Short single stranded overhangs at each end
- Can stick DNA molecule back together
7
Q
DNA cloning steps
A
- Enzyme cuts animal DNA
- ANimal gene injected into bacteria plasmid
- Recombinant plasmid inserted inside bacterium
- Replication inside the bacteria
- Many copies of recombinant plasmid produced - can produce the protein that we’re looking for
8
Q
What can be used as cloning vectors
A
- Plasmids
- Bacteriophage
- Cosmids
- Yeast artificial chromosomes (for v large DNA)
