Recombinant DNA Assignment

Recombinant DNA Assignment
Module 9C Recombinant DNA Study guide and Extra credit
Clinical Genetics
10 Possible points
Must be turned in by deadline for credit
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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