Restiction Enzymes: How is DNA Manipulated? Teacher's Guide & Key

This assignment is usually given with the unit on biotechnology: Biotechnology Notes and Powerpoint Presentation

Related Assignments on biotechnology

Who Ate the Cheese - use restriction enzymes to cut DNA and place on a chart to simulate movement of fragments during eletrophoresis

Recombinant DNA simulation - cut human and bacteria DNA at specific points using restriction enzymes, splice together to make rDNA

Gel Electrophoresis Lab - using a kit, students practice loading wells and analyzing DNA to detect a specific gene

                    

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.


BAMHI

G / G A T C C

PstI

C T G C A / G

HindIII

A / A G C T T

HhaI

G C G / C

EcoRI

G / A A T T C

HpaII

C / C G G

SalI

G / T C G A C

 

 

 

HindII

G T C / G A C
C A G / C T G

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. This is actually tedious to accomplish, I've found it easier if you actually cut the enzymes from the above chart and use them to find the corresponding locations on the sequences below, you may also wish to only assign the first couple because it takes so long to find the matches.


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'

Enzyme: ___EcoRI____

2.

5' G T G G / G A T C C C T T A 3'
3' C A C C C T A G / G G A A T 5'

Enzyme: ___BAMHI________

3.

5' A C G C C T C / C G G A G A 3'
3' T G C G G A G G C / C T C T 5'

Enzyme: ____HpaII____

4.

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'

Enzyme: __HindIII________

5.

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'

Enzyme: ____HhaI________

3.  Create a restriction enzyme that will remove the gene of interst.  Give it a name too!

enzyme

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 make copies of the gene___
d) What are the potential applications for making a new flu? ______developing vaccines___________