Download Recombinant DNA WS

Making Recombinant DNA
Name
Per.
Part A.
1. Sequence each event of creating protein using recombinant DNA in the correct order. Use the
word bank to fill in the blanks in the sequencing cutouts.
2. Draw a picture to match the process, making sure to label your drawing with the underlined
words from the cut-outs. Use a different color to represent each of the following: gene of interest, plasmid,
bacterium.
Word Bank. Words may be repeated.
gene
DNA ligase
restriction site
ligation
sticky ends
transformation
recognition sequence
recombinant plasmid
Event
1.
restriction digest
restriction enzyme
Drawing
A.
2.
3.
B.
4.
5.
C.
6.
7.
D.
8.
Part B. Get 3 small pieces of blue tape from your teacher. Identify/show the following terms using the DNA
and plasmid cutouts given to you:
I got it! Stamp
ligation
recognition sequence
restriction digest
sticky ends
recombinant plasmid
restriction enzyme
Part C. Identify the following for each of the restriction enzymes.
1. Hind III: 5’…AAGCTT …3’
3’…T TCGAA…5’
a. Recognition sequence:
b. Sticky ends:
2. BamH I: 5’…GGATCC…3’
3’…CCTAGG…5’
a. Recognition sequence:
b. Sticky ends:
a. Recognition sequence:
b. Sticky ends:
a. Recognition sequence:
b. Sticky ends:
3. EcoRI:
5’…GAATTC…3’
3’…CTTAAG…5’
4. 5’…TGGCCA…3’
3’…ACCGGT…5’
5. a. What is the GFP gene?
b. Where does the GFP gene come from?
c. Which restriction enzyme is used to cut the GFP gene with?
d. What sticky ends will the GFP have?
Part D. Answer the questions regarding the Plasmid P.
Fragment Y
1. What is the base pair length of the plasmid?
2. Gene A (750 bp) will be cut with Enzyme B and inserted into Plasmid P.
Plasmid P
8350 bp
Enzyme A
1425 bp
Fragment X
a. How many pieces of plasmid will result from the cut?
b. What is the length of each plasmid piece?
c. The cut plasmid is run through a gel. Draw & label each band(s) on the
resulting gel. The loading well has been drawn for you.
d. What is the resulting base pair length after insertion?
Show work.
3. Gene B (1450 bp) will be cut with Enzyme A and Enzyme B, and inserted into Plasmid P.
a. How many pieces of plasmid will result from the cut?
b. What is the length of each plasmid piece?
c. The cut plasmid is run through a gel. Draw & label each band(s) on the resulting gel.
d. What is the resulting base pair length when inserted with Gene B? Show work.
Enzyme B
2130 bp
the plasmid is cut at the _______________ in the process called _________________________
_____________________ takes place when the bacteria accepts the plasmid with the gene
the gene is inserted into the plasmid when the enzyme, ___________, joins the ________________ together in the process called __________
the __________ of interest is isolated
this plasmid with the gene of interest inserted is called the ___________________
the bacteria multiplies and amplifies the _____________
the ends of the cut DNA forms are called _________________________
a ____________________ cuts the gene at the ___________ by finding the ____________________
the plasmid is cut at the _______________ in the process called _________________________
_____________________ takes place when the bacteria accepts the plasmid with the gene
the gene is inserted into the plasmid when the enzyme, ___________, joins the ________________ together in the process called __________
the __________ of interest is isolated
this plasmid with the gene of interest inserted is called the ___________________
the bacteria multiplies and amplifies the _____________
the ends of the cut DNA forms are called _________________________
a ____________________ cuts gene at the ___________ by finding the ____________________