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E. coli insulin factory lab

Day 1

Newbie

 

Do Now

Read the Introduction of the E. Coli Insulin Factory Lab. [Click the hyperlink to access the lab notebook) Answer the following questions.

  1. What is a plasmid?

  2. Define Recombinant DNA

  3. What is the function of restriction enzymes?

 

Using the image at the top of the lab, state the steps to creating recombinant DNA. Use the following terms to write out the 4 steps needed to create recombinant DNA in bacteria:

  • restriction enzyme

  • plasmid

  • gene of interest

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Semi-Pro

 

Watch a short clip video on Diabetes and the importance of Insulin production. Answer a series of questions based on the video

Access the worksheet to answer the questions based on the video.

 

 

 

 

Read the procedures of the lab beginning on page 2 of the lab step 6. Steps 1-5 have been completed for you. An alternative to this is watching a video of the procedure. Begin the video at 3:48.

 

 

 

  • Look at your plasmid and your DNA strip. What differences do you notice between the two?

 

  • Identify what antibiotic resistance your plasmid has by circling the resistance genes in your lab on page 1 of the lab. Label your resistance gene on your plasmid.

 

  • Use the restriction enzymes to locate their positions on the plasmid. You can create a surrounding box around the sequence and label it to help identify the restriction enzyme sequence. Complete the restriction map in your lab book identifying approximate locations of each restriction enzyme on your plasmid. You should have at least 8 sites marked mapping the restriction enzyme sites, and one for each antibiotic resistant gene found on your plasmid.

 

  • Using the restriction enzymes determine which restriction enzyme can be used to cut the plasmid ONCE and cut out the insulin gene from the DNA strand TWICE (at the beginning and end of the insulin)!!! Remember not to cut any part of the insulin gene. Find the restriction enzyme code in both the plasmid and the DNA strand. Record how many times the enzyme can cut on both the plasmid and the DNA strand on your data table.

 

  • Work on completing the data table that indicates which restriction enzymes can be used to cut the plasmid and the strand of DNA. You must explain why you can or cannot use the restriction enzymes that are provided.

 

TASKS TO COMPLETE THIS PERIOD

Identify antibiotic resistance gene(s) on your plasmid

Restriction map

Begin working on Restriction enzyme data sheet

Day 2

Newbie

 

Do Now

Look at the image of the plasmid map. Answer the following question using the map:

 

 

 

 

 

 

 

 

 

 

 

 

  1. How many pieces will exist if restriction enzyme EcoR I cuts the plasmid?

  2. How many pieces will exist if restriction enzyme Nco I cuts the plasmid?

  3. How many pieces will exist if restriction enzyme BamH I cuts the plasmid?

 

Semi-Pro

Watch a short clip video on Antibiotic Resistance on Bacteria. Answer a series of questions based on the video. Access the worksheet to answer the questions based on the video. The questions are on page 2 of this document.

 

 

 

 

Continue working on the lab.

  • Using the restriction enzymes determine which restriction enzyme can be used to cut the plasmid ONCE and cut out the insulin gene from the DNA strand TWICE (at the beginning and end of the insulin)!!! Remember not to cut any part of the insulin gene. Find the restriction enzyme code in both the plasmid and the DNA strand. Record how many times the enzyme can cut on both the plasmid and the DNA strand on your data table.

 

  • Complete the data table that indicates which restriction enzymes can be used to cut the plasmid and the strand of DNA. You must explain why you can or cannot use the restriction enzymes that are provided.

 

  • Cut the insulin gene out of the strand of DNA. Cut open the plasmid. Create recombinant DNA by taping together the sticky ends of the DNA. Write all group members names on the inside of the completed recombinant DNA.

 

 

 

Boss Level

 

Genetically Modified Organisms are the new wave in technology for biologists. This innovative technology has brought about many solutions to many problems. Because these technologies are so new they leave many unanswered questions. Provide evidence that supports the PRO’s or CON’s of GMO’s.

 

Write your defense in PEA (Perspective Evidence Analysis) format.

Perspective: State your case. GMO's are beneficial.... or.... GMO's are damaging

Evidence: What scientific proof do you have that supports your analysis? You should include at least 3 peices of evidence that support your perspective.

Analysis: Why is this evidence useful to support your perspective? What evidence is their that disapproves the other perspective?

 

You will need to do some research to provide credible evidence to support your case.  Include references of all sources you used.

 

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