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Suppose we want to insert the human insulin gene into E. coli using pBR322 as the expression vector. 1. Explain how you will identify colonies that have been successfully transformed. 2. Identify all restriction enzyme(s) that would be appropriate for inserting the gene into pBR322. 3. Explain why the enzyme(s) from #2 are appropriate. Think about the function of the sequences in the expression vector when you answer this question. 4. The plasmid map shows that amp and ori are oriented in one direction and tet is in the opposite direction. How is this possible? 5. Suppose you decide to run the DNA on a gel to check each step. Draw the gel that would result from the following: Lane 1 molecular weight marker Lane 2 undigested pBR322 Lane 3 pBR322 digested with your selected restriction enzyme (identify which enzyme) Lane 4 pBR322 digested with your selected restriction enzyme then ligated with human insulin gene

Suppose we want to insert the human insulin gene into E. coli using pBR322 as the expression vector. 1. Explain how you will identify colonies that have been successfully transformed. 2. Identify all restriction enzyme(s) that would be appropriate for inserting the gene into pBR322. 3. Explain why the enzyme(s) from #2 are appropriate. Think about the function of the sequences in the expression vector when you answer this question. 4. The plasmid map shows that amp and ori are oriented in one direction and tet is in the opposite direction. How is this possible? 5. Suppose you decide to run the DNA on a gel to check each step. Draw the gel that would result from the following: Lane 1 molecular weight marker Lane 2 undigested pBR322 Lane 3 pBR322 digested with your selected restriction enzyme (identify which enzyme) Lane 4 pBR322 digested with your selected restriction enzyme then ligated with human insulin gene

Human Anatomy & Physiology (11th Edition)
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
Section: Chapter Questions
Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
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Suppose we want to insert the human insulin gene into E. coli using pBR322 as the expression vector. 1. Explain how you will identify colonies that have been successfully transformed. 2. Identify all restriction enzyme(s) that would be appropriate for inserting the gene into pBR322. 3. Explain why the enzyme(s) from #2 are appropriate. Think about the function of the sequences in the expression vector when you answer this question. 4. The plasmid map shows that amp and ori are oriented in one direction and tet is in the opposite direction. How is this possible? 5. Suppose you decide to run the DNA on a gel to check each step. Draw the gel that would result from the following: Lane 1 molecular weight marker Lane 2 undigested pBR322 Lane 3 pBR322 digested with your selected restriction enzyme (identify which enzyme) Lane 4 pBR322 digested with your selected restriction enzyme then ligated with human insulin gene
1. Explain how you will identify colonies that have been successfully transformed. 2. Identify all restriction enzyme(s) that would be appropriate for inserting the gene into pBR322. 3. Explain why the enzyme(s) from #2 are appropriate. Think about the function of the sequences in the expression vector when you answer this question. 4. The plasmid map shows that amp and ori are oriented in one direction and tet is in the opposite direction. How is this possible? 5. Suppose you decide to run the DNA on a gel to check each step. Draw the gel that would result from the following: Lane 1 molecular weight marker Lane 2 undigested pBR322 Lane 3 PBR322 digested with your selected restriction enzyme (identify which enzyme) Lane 4 PBR322 digested with your selected restriction enzyme then ligated with human insulin gene
Transcribed Image Text:1. Explain how you will identify colonies that have been successfully transformed. 2. Identify all restriction enzyme(s) that would be appropriate for inserting the gene into pBR322. 3. Explain why the enzyme(s) from #2 are appropriate. Think about the function of the sequences in the expression vector when you answer this question. 4. The plasmid map shows that amp and ori are oriented in one direction and tet is in the opposite direction. How is this possible? 5. Suppose you decide to run the DNA on a gel to check each step. Draw the gel that would result from the following: Lane 1 molecular weight marker Lane 2 undigested pBR322 Lane 3 PBR322 digested with your selected restriction enzyme (identify which enzyme) Lane 4 PBR322 digested with your selected restriction enzyme then ligated with human insulin gene
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