Recombinant DNA
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Module9CHomeworkextracredit.pptxModule 9C Recombinant DNA Study guide and Extra credit
Clinical Genetics
10 Possible points
Must be turned in by deadline for credit
1
This is Extra credit homework Create it for your own study guide
What is the cloning advantage to have a polylinker in the vector?
___ (1 pt)
Why is transformed E.coli resistant to ampicillin?
___ (1 pt)
Do the drawings and answer the questions posed on the next slides
Procedure for cloning a recombinant protein
Combine measured amounts of the DNA insert with the vector, and add ligase. Incubate. Sticky ends align and ligase sews it up
Draw a picture of the new plasmid, using different colors for the vector and the DNA insert. (1 pt)
Most of the original plasmid will reform with itself. Draw that. Make a notation of the ampicillin resistance gene. (1 pt)
Procedure for cloning a recombinant protein
Transform the recombinant plasmid mixture into competent E.coli. And plate the mixture on a plate with Ampicillin
If nothing grows on that plate, there can be 2 causes. What are they? (1 pt) __
How can you tell which was the cause (what control should you run)? (1 pt) __
What happens to the LacZ gene?
Draw the pBluescript plasmid, and highlight the LacZ gene in blue.
Redraw the plasmid with a DNA insert that is 500 bp long. Be sure to change the LacZ gene in your drawing. 1 pt for the 2 pictures in your answer.
LacZ (2 pts for this slide)
What enzyme is produced by the LacZ gene?
What does that enzyme do to X-gal?
Which colonies in the figure have plasmid transformants?
Which colonies have the DNA insert?
Vectors (1 pt for this slide)
Name 2 limitations to using plasmid vectors and E. coli cloning
__
__
What type of vector can replicate in E. coli and other species as well?
__
Highlight the DNA from species 1 in yellow. Teach the slide sequence to someone outside of class (no points)
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