How is DNA Manipulated?
1. Explain the following terms and their role in recombinant DNA technology
a) Restriction enzyme ___cuts DNA into fragments_____
b) Recognition site ___specific site or spot where the enzyme cuts__
c) Sticky end ______area of DNA where bases are ready to be paired, they will "stick" to
matching DNA__
d) Ligation _____________the joining of two different DNA________________
e) rDNA ________the new DNA is called DNA, or recombinant DNA_____________
Here are some restriction enzymes and their sites.
G/GATCC
PstI
CTGCA/G
HindIII
A/AGCTT
HhaI
GCG/C
EcoRI
G/AATTC
HpaII
C/CGG
SalI
G/TCGAC
HindII
GTC/GAC
BAMHI
CAG/CTG
2. On each of the sequences below, determine which restriction enzyme could be used to
splice the DNA and indicate where the cut will be made and the enzyme used.
1.
5' T T T ​G / A A T T C​ A G A T 3'
3' A A A ​C T T A A / G​ T C T A 5'
2.
3.
Enzyme:
3' C A C ​C C T A G / G​ G A A T 5'
_
5' A C G C C T​ C / C G G​ A G A 3'
5' T T ​A / A G C T T​ A A G A A G C T T 3'
3' A A ​T T C G A / A​ T T C T T C G A A 5'
5.
___​EcoRI​____
5' G T G ​G / G A T C C​ C T T A 3'
3' T G C G G A ​G G C / C​ T C T 5'
4.
Enzyme:
5' A A ​G C G / C​ G T C G A C T T A T A 3'
3' T T ​C / G C G​ C A G C T G A A T A T 5'
___​BAMHI​_______
Enzyme:
____​HpaII​____
Enzyme:
__​HindIII​________
Enzyme:
____​HhaI​________
3. Create a restriction enzyme that will remove the gene of interst. Give it a name too!
4. The following DNA sequence is from a virus that is dangerous, scientists want to use a
restriction enzyme to cut the virus into bits. They do not need sticky ends because the do not
plan to combine it with other DNA. Use HindII to show how this DNA would be cut. How many
pieces would you have? __​4​__
G A A A A ​G T C / G A C ​A A G G C A ​G T C / G A C​ T T T T A A A A​ G T C / G A C​ A T G C
C T T T T ​C A G / C T G​ T T C C G T ​C A G / C T G​ A A A A T T T T ​C A G / C T G​ T A C G
How is the Recombinant DNA (rDNA) Used?
a) Which genes are being isolated from the flu virus? ____​HA
and NA​_____
b) Where are the flu genes (HA and NA) placed? ____​plasmids​___________
c) Where do plasmids come from and why are they imporant in this process? _​they
copies of the gene​___
d) What are the potential applications for making a new flu? ______​developing
vaccines​___________
make